Hi there GPCR enthusiasts! Join us this week for a deep dive into the world of GPCRs. This week's GPCR papers feature publications from Drs. Ines Liebscher, Ross Cheloha and Silvio Gutkind. Before we dive into this week’s news, we’d like to draw your attention to some of our events and resources: Please mark your calendars for July 21st for our next Dr. GPCR Symposium on Structural and Molecular Insights on GPCR Activation. Our website is updated as we confirm speakers so please check back often. We also seek speakers and poster presenters for the next symposium and beyond! Showcase your work by emailing us at Hello@DrGPCR.com. Stay updated with Dr. GPCR News by subscribing to our monthly newsletter. Our founder Dr. Yamina A. Berchiche is at the Molecular Pharmacology GRC, if you are also on-site, please stop by and say hi. And now let’s dive into the weekly Classified GPCR News at a glance for June 5th to June 11th, 2023. Adhesion GPCRs Collagen VI Is a Gi-Biased Ligand of the Adhesion GPCR GPR126/ADGRG6. Hexahydroquinoline Derivatives are Selective Agonists for the Adhesion G Protein-Coupled Receptor ADGRG1/GPR56. GPCR Activation and Signaling Altered Signaling and Desensitization Responses in PTH1R Mutants Associated with Eiken Syndrome Antagonistic interactions between odorants alter human odor perception. Autocrine Proteinase-Activated Receptor (PAR) signaling in PC3 prostate cancer cells. Statin-induced increase in actin polymerization modulates GPCR dynamics and compartmentalization. Class B1 GPCR activation by an intracellular agonist. The G protein-coupled receptor neuropeptide receptor-15 modulates larval development via the transforming growth factor-β DAF-7 protein in Caenorhabditis elegans. GPCR Binders, Drugs, and more Adenosine A2A Receptor (A2AAR) Ligand Screening Using the 19F-NMR Probe FPPA. GPCRs in Cardiology, Endocrinology, and Taste Discovery of paralogous GnRH and corazonin signaling systems in an invertebrate chordate. GPCRs in Neuroscience Early pheromone perception remodels neurodevelopment and accelerates neurodegeneration in adult C. elegans. Methods & Updates in GPCR Research A Peptidisc-based Survey of the Plasma Membrane Proteome of a Mammalian Cell. Reviews, GPCRs, and more Diversification processes of teleost intron-less opsin genes. Regulation of the length of neuronal primary cilia and its potential effects on signalling. Structural and Molecular Insights into GPCR Function Structural insights into ligand recognition and activation of the medium-chain fatty acid-sensing receptor GPR84. Stabilization of pre-existing neurotensin receptor conformational states by β-arrestin-1 and the biased allosteric modulator ML314. GPCRs in Immunology and Oncology The GPCR–Gαs–PKA signaling axis promotes T cell dysfunction and cancer immunotherapy failure Industry News X4 Pharmaceuticals Expected to Join Russell 3000® Index. Novartis bolsters innovative medicines strategy and renal pipeline with agreement to acquire Chinook Therapeutics for USD 3.2bn upfront (USD 40 / share). EyeBio Announces First Patients Dosed in Phase 1b/2 Trial of Restoret™ in Neovascular Age Related Macular Degeneration and Diabetic Macular Edema. Confo Therapeutics Enters Into Research Collaboration For GPCR-Targeting Antibody Discovery With AbCellera. GPCR Events, Meetings, and Webinars GPCRs in drug discovery: challenges & solutions (June 19 - 23, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Retreat (November 2 - 4, 2023) NEW Structure, Mechanism, and Drug Interactions of GPCRs, Ion Channels, and Transport Proteins (March 24 - 29, 2024) GPCR Jobs NEW Scientist I / Scientist II DOE (Native Mass Spectrometry) Senior Vice President of Global Medical Affairs Postdoctoral Position on Gliovascular Unit of Glioblastoma Associate Director / Director, Clinical Operations Postdoctoral Research Associate Lab Manager Explore Dr. GPCR Ecosystem

Hello GPCR friends👋, Stay informed with the latest developments in GPCR research and industry news! Happy reading📚! This week, we would like to highlight two new publications: Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics. Tagged for arrest: 'Barcode' determines receptor's fate. Our next Dr. GPCR Symposium is coming up on July 21st on Structural and Molecular Insights on GPCR Activation. We are looking for speakers and poster presenters! If you'd like to showcase your work, please email us at Hello@DrGPCR.com. For more information, follow the Dr. GPCR News by subscribing to our monthly newsletter. We look forward to seeing you in a few days at the Molecular Pharmacology GRC and the associated Molecular Pharmacology GRS in Switzerland. We would also like to take a moment to thank the organizers for inviting Dr. Yamina Berchiche, our founder. Below is your Classified GPCR News at a glance for May 29th to June 4th, 2023. GPCR Activation and Signaling Therapeutic antagonism of the neurokinin 1 receptor in endosomes provides sustained pain relief. Molecular mechanism of fatty acid activation of FFAR1. Conformational flexibility of β-arrestins - How these scaffolding proteins guide and transform the functionality of GPCRs. Gβγ signaling regulates microtubule-dependent control of Golgi integrity. Ligand-dependent intracellular trafficking of the G protein-coupled P2Y6 receptor. GPCR Binders, Drugs, and more ACE2 Activation by Tripeptide IRW (Ile-Arg-Trp) Depends on the G Protein-Coupled Receptor 30 Signaling Cascade. Molecular insights into peptide agonist engagement with the PTH receptor. An Atomic Level Investigation of Sodium Ions Regulating Agonist and Antagonist Binding in the Active Site of a Novel Target 5HT2BR Against Drug-Resistant Epilepsy. GPCRs in Cardiology, Endocrinology, and Taste GPR97 deficiency ameliorates renal interstitial fibrosis in mouse hypertensive nephropathy. Bitter Odorants and Odorous Bitters: Toxicity and Human TAS2R Targets. GPCRs in Neuroscience DRD1 signaling modulates TrkB turnover and BDNF sensitivity in direct pathway striatal medium spiny neurons. The contribution of endocytosis to sensitization of nociceptors and synaptic transmission in nociceptive circuits. The crosstalk between 5-HT2AR and mGluR2 in schizophrenia. GPCRs in Oncology and Immunology A2aR on lung adenocarcinoma cells: A novel target for cancer therapy via recruiting and regulating tumor-associated macrophages. Coronavirus Porcine Epidemic Diarrhea Virus Utilizes Chemokine Interleukin-8 to Facilitate Viral Replication by Regulating Ca2+ Flux. CircFKBP5 Suppresses Apoptosis and Inflammation and Promotes Osteogenic Differentiation. Transcriptomic analysis of human cytomegalovirus to survey the indirect effects on renal transplant recipients. Pharmacological inhibition of neuropeptide Y receptors Y1 and Y5 reduces hypoxic breast cancer migration, proliferation, and signaling. Methods & Updates in GPCR Research Illuminating the brain-genetically encoded single wavelength fluorescent biosensors to unravel neurotransmitter dynamics. Activity Models of Key GPCR Families in the Central Nervous System: A Tool for Many Purposes. Structure-based pharmacophore modeling 1. Automated random pharmacophore model generation. NMR applications to GPCR recognition by peptide ligands. How can we improve the measurement of receptor signaling bias? Reviews, GPCRs, and more Advancements in G protein-coupled receptor biosensors to study GPCR-G protein coupling. Application of computational methods for class A GPCR Ligand discovery. New paradigms in purinergic receptor ligand discovery. Targeting biased signaling by PAR1: function and molecular mechanism of parmodulins. Lysophosphatidic acid, a simple phospholipid with myriad functions. Structural and Molecular Insights into GPCR Function Pharmacological hallmarks of allostery at the M4 muscarinic receptor elucidated through structure and dynamics. The role of G protein conformation in receptor-G protein selectivity. Components of TOR and MAP kinase signaling control chemotropism and pathogenicity in the fungal pathogen Verticillium dahliae. New insights into GPCR coupling and dimerisation from cryo-EM structures. Computational insights into ligand-induced G protein and β-arrestin signaling of the dopamine D1 receptor. Understanding the Rhodopsin Worldview Through Atomic Force Microscopy (AFM): Structure, Stability, and Activity Studies. Scribble scrambles parathyroid hormone receptor interactions to regulate phosphate and vitamin D homeostasis. Industry News Structure Therapeutics Announces Poster Presentations of its Oral GLP-1 Receptor Agonist GSBR-1290 at the American Diabetes Association 83rd Scientific Sessions Domain Therapeutics' Nomination of DT-9045: A Groundbreaking NAM for Immunooncology Structure Therapeutics to Present at Jefferies Healthcare Conference Tagged for arrest: “Barcode” determines receptor’s fate. GPCR Events, Meetings, and Webinars FREE Symposium - IPI Surfacing (June 15, 2023) GPCRs in drug discovery: challenges & solutions (June 19 - 23, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Retreat (November 2 - 4, 2023) GPCR Jobs NEW Senior Vice President of Global Medical Affairs NEW Postdoctoral Position on Gliovascular Unit of Glioblastoma Associate Director / Director, Clinical Operations Postdoctoral Research Associate Lab Manager Vice President, Medical Affairs Explore Dr. GPCR Ecosystem

Hi GPCR lovers👋, Ready for exciting news in the GPCR world? Read them below! You can now watch the live talk recordings from the previous symposiums, check them out here. Also, let's congratulate Lauri Urvas on his first first-author paper! Check out GPCR papers featuring Drs. Bryan Roth, Peter Gmeiner, and Thomas P. Sakmar this week. For Dr. GPCR News, please subscribe to our monthly newsletter. Below is your Classified GPCR News at a glance for May 22 to 28, 2023. GPCR Activation and Signaling The adhesion GPCRs CELSR1-3 and LPHN3 engage G proteins via distinct activation mechanisms. Multiple Subthreshold GPCR Signals Combined by the G-Proteins Gαq and Gαs Activate the Caenorhabditis elegans Egg-Laying Muscles. Modulating GPCR and 14-3-3 protein interactions: prospects for CNS drug discovery. GPCR Binders, Drugs, and more Design of Drug Efficacy Guided by Free Energy Simulations of the β2 -Adrenoceptor. Bioorthogonal Tethering Enhances Drug Fragment Affinity for G Protein-Coupled Receptors in Live Cells. Differential Responses of the GLP-1 and GLP-2 Receptors to N-Terminal Modification of a Dual Agonist. GPCRs in Cardiology, Endocrinology, and Taste Pathophysiology and pharmacology of G protein-coupled receptors in the heart. GPCRs in Neuroscience PACAP key interactions with PAC1, VPAC1, and VPAC2 identified by molecular dynamics simulations. Therapeutic antagonism of the neurokinin 1 receptor in endosomes provides sustained pain relief. Structural and Molecular Insights into GPCR Function Cholesterol Biases the Conformational Landscape of the Chemokine Receptor CCR3: A MAS SSNMR-Filtered Molecular Dynamics Study. The intertwining roles of caveolin, oxytocin receptor, and the associated signalling pathways in prostate cancer progression. A key GPCR phosphorylation motif discovered in arrestin2⋅CCR5 phosphopeptide complexes. Divergent regulation of α-arrestin ARRDC3 function by ubiquitination. Molecular mechanism of fatty acid activation of FFAR1. Structural genomics of the human dopamine receptor system Structural Insights into Molecular Recognition and Receptor Activation in Chemokine–Chemokine Receptor Complexes Industry News Mavorixafor reduces infection rates among patients with WHIM syndrome Bristol Myers Squibb’s Investigational LPA1 Antagonist Reduces the Rate of Lung Function Decline in Patients with Idiopathic Pulmonary Fibrosis Exscientia Business Update for First Quarter 2023 Structure Therapeutics Initiates Phase 2a Study of Oral GLP-1 agonist GSBR-1290 for the Treatment of Type 2 Diabetes and Obesity Prof. Andrew Hopkins, Exscientia's CEO, will speak at the University of Chicago's Distinguished Seminar Series on Population and Precision Health next week. Your body naturally produces opioids without causing addiction or overdose – studying how this process works could help reduce the side effects of opioid drugs GPCR Events, Meetings, and Webinars The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) NEW GPCRs in drug discovery: challenges & solutions (June 19 - 23, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) NEW GPCR Retreat (November 2 - 4, 2023) GPCR Jobs NEW Associate Director / Director, Clinical Operations Postdoctoral Research Associate Lab Manager Vice President, Medical Affairs Associate Scientist - Technology Explore Dr. GPCR Ecosystem

At the cellular level, the plasmatic membrane is a thin layer of lipids that surrounds the entire cell allowing the differentiation between the intracellular and extracellular environment. The physical barrier this lipid bilayer creates is dynamic and interactive, becoming the foundation for many interactions involved in GPCR signaling6. The cellular membrane's composition, organization, and physical properties might impact ligand binding, receptor activation, G protein interactions, signaling dynamics, and signal termination, highlighting the importance of studying their role in the activation of GPCRs2. β-arrestins are cytosolic proteins that translocate to the plasma membrane upon GPCR activation, then regulate trigger receptor internalization via interaction with the adaptor protein 2 (AP2) and clathrin heavy chain mediating G protein-independent effects. At this point, the lipid bilayer serves as a platform for the membrane recruitment of β-arrestins. Lipid molecules, such as phosphoinositides, can bind to specific domains of β-arrestins, promoting their association with the plasma membrane. Therefore the lipid composition of the bilayer can influence the kinetics and efficiency of β-arrestin recruitment to the membrane, regulating their interactions with activated GPCRs2-6. Understanding the interplay between GPCRs and β-arrestins and how this complex operates on the plasma membrane of living cells was the goal achieved by Jak Grimes et al. Based on an ingenious combination of multicolor single-molecule microscopy approach with molecular dynamics simulations, they dissected the sequence of events in receptor-β-arrestin interactions at the plasma membrane of living cells with ~20 nm spatial and ~30 ms temporal resolution. Whit this scope, the authors challenged the current model, wich suggest that β-arrestin translocates from the cytosol to bind an active receptor on the plasma membrane directly and remains attached to the same receptor until they reach clathrin-coated pits (CCPs)1. However, the evidence obtained in this study proposes novel molecular mechanisms in which β-arrestin exhibits spontaneous pre-association with the plasma membrane. This pre-association enables β-arrestin to explore its surroundings through lateral diffusion and engage in highly transient interactions with receptors, ultimately leading to β-arrestin activation. This event extends the duration of β-arrestin at the plasma membrane, enabling it to independently reach clathrin-coated pits (CCPs) without solely relying on the initial and short-lived receptor-β-arrestin complexes 1. Based on all the information collected, the authors proposed the following multistep model for receptor-β-arrestin interactions under unstimulated and stimulated conditions: Unstimulated condition: Inactive β-arrestin in the cytosol spontaneously binds to the plasma membrane by inserting the C-edge into the lipid bilayer, allowing it to explore space via lateral diffusion. Most β-arrestin molecules remain on the plasma membrane briefly before dissociating and returning to the cytosol. Stimulated condition in the presence of a stimulated receptor: Spontaneous insertion into the plasma membrane β-arrestin. β-arrestin reaches the receptor via lateral. Transient interaction with the receptor catalyzes β-arrestin activation, including β-arrestin inter-domain rotation and extension of the finger loop. Following dissociation from the receptor, the interaction of the extended finger loop with the lipid bilayer likely contributes to stabilizing β-arrestin in a membrane-bound, active-like conformation. The above causes β-arrestin molecules to stay longer and accumulate on the plasma membrane, allowing them to reach CCPs vial lateral diffusion separately from the activating receptors. The increase in active β-arrestin molecules and the time they spend diffusing on the plasma membrane leads to their recruitment and accumulation in CCPs via interaction with AP2 and clathrin. β-arrestin molecules tethered to CCPs bind receptors diffusing on the plasma membrane, also causing their recruitment and accumulation in CCPs. Although this study has limitations, such as the absence of the flexible distal C-tail of βArr2 in the model used, its findings redefine the existing model of receptor-β-arrestin interactions. They shed light on the essential role of β-arrestin binding to the lipid bilayer for efficient interaction between β-arrestin and the receptor. Read the complete article here: https://www.ecosystem.drgpcr.com/gpcr-binders-drugs-and-more/in-depth-molecular-profiling-of-an-intronic-gnao1-mutant-as-the-basis-for-personalized-high-throughput-drug-screening References Grimes, J., Koszegi, Z., Lanoiselée, Y., Miljus, T., O'Brien, S. L., Stepniewski, T. M., Medel-Lacruz, B., Baidya, M., Makarova, M., Mistry, R., Goulding, J., Drube, J., Hoffmann, C., Owen, D. M., Shukla, A. K., Selent, J., Hill, S. J., & Calebiro, D. (2023). Plasma membrane preassociation drives β-arrestin coupling to receptors and activation. Cell, 186(10), 2238–2255.e20. https://doi.org/10.1016/j.cell.2023.04.018 Janetzko, J., Kise, R., Barsi-Rhyne, B., Siepe, D. H., Heydenreich, F. M., Kawakami, K., Masureel, M., Maeda, S., Garcia, K. C., von Zastrow, M., Inoue, A., & Kobilka, B. K. (2022). Membrane phosphoinositides regulate GPCR-β-arrestin complex assembly and dynamics. Cell, 185(24), 4560–4573.e19. https://doi.org/10.1016/j.cell.2022.10.018 Latorraca, N.R., Wang, J.K., Bauer, B., Townshend, R.J.L., Hollingsworth, S.A., Olivieri, J.E., Xu, H.E., Sommer, M.E., and Dror, R.O. (2018). Molecular mechanism of GPCR-mediated arrestin activation. Nature 557, 452–456. https://doi.org/10.1038/s41586-018-0077-3 Pierce, K.L., and Lefkowitz, R.J. (2001). Classical and new roles of b-arrestins in the regulation of G-protein-coupled receptors. Nat. Rev. Neurosci. 2, 727–733. https://doi.org/10.1038/35094577. Reiter, E., Ahn, S., Shukla, A.K., and Lefkowitz, R.J. (2012). Molecular mechanism of b-arrestin-biased agonism at seven-transmembrane receptors. Annu. Rev. Pharmacol. Toxicol. 52, 179–197. https://doi.org/10. 1146/annurev.pharmtox.010909.105800 Tsao, P. I., & von Zastrow, M. (2001). Diversity and specificity in the regulated endocytic membrane trafficking of G-protein-coupled receptors. Pharmacology & therapeutics, 89(2), 139–147. https://doi.org/10.1016/s0163-7258(00)00107-8

Hi GPCR buffs 👋! This week's GPCR papers feature a publication from Dr. Marta Filizola. Check it out! The Dr. GPCR Symposium on GPCR Activation and Signaling was a great success. We express our gratitude to the speakers for their excellent talks. Our next symposium is on July 21st, 2023, focusing on Structural and Molecular Insights into GPCR Activation. If you are interested in giving a talk, contact us at Hello@DrGPCR.com, and if you would like to present a poster, submit this form here. Take the chance to contribute to this exciting event! For Dr. GPCR News, please subscribe to your monthly newsletter. Below is your Classified GPCR News at a glance for May 15th to 21st, 2023. GPCR Activation and Signaling Monitoring the Reversibility of GPCR Signaling by Combining Photochromic Ligands with Label-free Impedance Analysis. FSLLRY-NH2, a protease-activated receptor 2 (PAR2) antagonist, activates mas-related G protein-coupled receptor C11 (MrgprC11) to induce scratching behaviors in mice. ACE2 Activation by Tripeptide IRW (Ile-Arg-Trp) Depends on the G Protein-Coupled Receptor 30 Signaling Cascade. Gαs slow conformational transition upon GTP binding and a novel Gαs regulator. GPCRs in Neuroscience Conformationally Selective 2-Aminotetralin Ligands Targeting the alpha2A- and alpha2C-Adrenergic Receptors. A vital role for PICK1 in the differential regulation of metabotropic glutamate receptor internalization and synaptic AMPA receptor endocytosis. GPCRs in Oncology and Immunology Stable Binding of Full-Length Chemerin is Driven by Negative Charges in the CMKLR1 N-terminus. Methods & Updates in GPCR Research Computational design of dynamic receptor-peptide signaling complexes applied to chemotaxis. Reviews, GPCRs, and more Analysis of the Dynamics of the Human Growth Hormone Secretagogue Receptor Reveals Insights into the Energy Landscape of the Molecule. Industry News X4 Pharmaceuticals Announces $65 Million Private Placement Priced At-the-Market Andrew Hopkins, Exscientia’s founder and CEO was elected a Fellow of the Academy of Medical Sciences Structure Therapeutics Announces Poster Presentations at the American Thoracic Society International Conference GPCR Events, Meetings, and Webinars SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) PEGS Boston (May 15 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs NEW Postdoctoral Research Associate NEW Lab Manager Vice President, Medical Affairs Associate Scientist - Technology Senior Research Scientist/Principal Scientist- Technology Tissue Culture Scientist/Tissue Culture Manager Postdoctoral Researcher Director - Targeted Oncology Explore Dr. GPCR Ecosystem

Hi GPCR lovers! Stay tuned for the latest buzz on GPCRs. This week's GPCR papers feature publications from Drs. Vladimir Katanaev and Patrick Sexton. Remember to join us tomorrow, May 19th, for the highly anticipated Dr. GPCR Symposium on GPCR Activation and Signaling! If you're interested in presenting a poster, there's still time. Simply submit your information using this form. To receive our monthly newsletter, please subscribe to Dr. GPCR News. Below is your Classified GPCR News at a glance for May 8th to 14th, 2023. GPCR Activation and Signaling Ligand-induced activation and G protein coupling of prostaglandin F2α receptor. Plasma membrane preassociation drives β-arrestin coupling to receptors and activation. Gβγ activates PIP2 hydrolysis by recruiting and orienting PLCβ on the membrane surface. GPCR Binders, Drugs, and more In-depth molecular profiling of an intronic GNAO1 mutant as the basis for personalized high-throughput drug screening. An allosteric modulator of the adenosine A1 receptor potentiates the antilipolytic effect in rat adipose tissue. GPCRs in Cardiology, Endocrinology, and Taste Molecular Mechanisms of PTH/PTHrP Class B GPCR Signaling and Pharmacological Implications. GPCRs in Oncology and Immunology DANGER Signals Activate G-Protein Receptor Kinases Suppressing Neutrophil Function and Predisposing to Infection After Tissue Trauma. Reviews, GPCRs, and more Recent Studies on Serotonin 5-HT2A Receptor Antagonists in Medicinal Chemistry: A Last Decades Survey. Mechanism of Activation of the Visual Receptor Rhodopsin. The application of artificial intelligence to accelerate GPCR drug discovery. Structural and Molecular Insights into GPCR Function New Insights into the Structure and Function of Class B1 GPCRs. Industry News InterAx Biotech to Apply its Leading AI and Cell Signalling Technology to Support Target Validation for a Sosei Heptares GPCR Discovery Program Salipro Biotech and DyNAbind Announce Milestone Achievement in Collaboration to Enable DEL for Membrane Proteins Professor Andrew Hopkins, Exscientia’s founder and CEO, was elected as a Fellow of the The Royal Society. Addex Announces ADX71149 Phase 2 Epilepsy Clinical Study’s Independent Interim Review Committee Recommends Continuing Study Addex Therapeutics in the 23rd BioEquity Europe Conference Simon Bekker-Jensen and Mette M Rosenkilde received two proof of concept grants from European Research Council (ERC) Addex Reports Q1 2023 Financial Results And Provides Corporate Update Structure Therapeutics Reports First Quarter 2023 Financial Results and Recent Highlights OMass Therapeutics presented at the 23rd BioEquity Conference British Patient Capital Invests £10m in next-generation drug development company OMass Therapeutics Sosei Heptares 2022 ESG report Exscientia Announces Sixth Molecule Created Through Generative AI Platform to Enter Clinical Stage X4 Pharmaceuticals Announces Positive Phase 3 Results Showing Mavorixafor Reduced the Rate, Severity, and Duration of Infections vs. Placebo in Participants Diagnosed with WHIM Syndrome GPCR Events, Meetings, and Webinars ASPET 2023 - American Society for Pharmacology and Experimental Therapeutics (May 18 - 21, 2023). SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) PEGS Boston (May 15 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs Vice President, Medical Affairs Associate Scientist - Technology Senior Research Scientist/Principal Scientist- Technology Tissue Culture Scientist/Tissue Culture Manager Postdoctoral Researcher Director - Targeted Oncology Senior Scientist - Antibody Engineering Scientist—Immuno-Oncology Convergent Research - Senior Scientist, Cell-Based Assay Development Explore Dr. GPCR Ecosystem

Artificial intelligence - Machine learning vs Deep Learning GPCRs have long been recognized as important drug targets, and numerous drugs that modulate GPCRs are already in clinical use for various diseases. The integration of AI in GPCR drug discovery has the potential to accelerate the identification and development of novel drugs, optimize drug design processes, and enhance the understanding of GPCR biology. High-throughput approaches used in drug discovery create large datasets regarding ligand synthesis and screening, ligand binding assays, signaling assays, cell imaging, protein structure determination, and omics applications, which can be analyzed by an established AI framework which involves three stages: 1) feature extraction or pattern identification; 2) vector space construction and metric definition where data is classified and compared; and 3) detection, prediction, or generation (e.g. prediction of a protein structure, or ligand design). Machine learning (ML) and deep learning (DL) are subfields of artificial intelligence (AI) that involve training algorithms to learn from data. While they share similarities, there are notable differences in respect to neural network architecture, complexity, data requirements, training time, computational resources and interpretability. ML algorithms typically use traditional ML models, such as decision trees and support vector machines which require handcrafted feature engineering, where domain experts manually select and engineer relevant features from the input data. While classical ML models are effective for datasets for which the relevant features are well understood, their use for datasets for which the relevant features of the input data are unknown, such as in drug discovery, is limited - the solution is DL. DL algorithms employ artificial neural networks with multiple layers of interconnected nodes which learn hierarchical representations of the data, eliminating the need for explicit feature engineering and allowing the exploitation of features that would not typically occur using classical ML algorithms. However, DL algorithms typically require large amounts of labelled training data comprising millions of parameters, becoming infeasible for a human to interpret, in contrast with alternative ML models which comprise handcrafted features and simpler models. For this reason, DL models generally require more computational resources (such as powerful GPUs) and longer training times. Artificial intelligence in GPCR drug discovery The use of AI in GPCR drug discovery has increased over the last decade and is revolutionising the way new GPCR-targeted drugs are developed. AI is providing a dramatic acceleration of the drug discovery process at multiple stages: 1. Classification: AI models can be used to distinguish GPCRs from non-GPCRs, and to classify GPCRs into families, subfamilies, sub-subfamilies, and subtypes. The models use a variety of data for the input, including amino acid sequences or structural data (from X-ray crystallography, cryoEM or molecular dynamics simulation experiments). 2. Mutations: ML methods can determine stabilising mutations that enable structure determination and can also predict the effect of mutagenesis on GPCR function. 3. Structure: the development of algorithms such DeepMind’s Alphafold2 (Jumper et al., 2021) and RoseTTAFold (Baek et al., 2021) allow the prediction of the 3D structure of a protein from the amino acid sequence even where no similar structure is known. 4. GPCR-ligand interactions: ML can predict GPCR-ligand interactions based on input data of protein sequences and molecular fingerprints coming from databases such as the GPCR-Ligand Association (GLASS), bindingDB and DrugBank. One major challenge is the identification of receptor subtype-selective ligands. In this context, BRS-3D was used to predict subtype-selective ligands for dopamine receptors and adenosine receptors (He, Ben, Kuang, Wang & Kong, 2016; Kuang, Feng, Hu, Wang, He & Kong, 2016). 5. Virtual screening: molecular docking and virtual screening can efficiently analyze large databases of compounds and predict their binding affinity to GPCRs, aiding the identification of lead compounds for further development. Paremeters such as ligand affinity for the receptor (pKi), the ability of a ligand to induce or inhibit a cellular response (pEC50 or pIC50, respectively), and how long the ligand remains bound to the receptor (koff), are used to short-list candidate drugs for further assessment. 6. De novo drug design: AI algorithms can generate new molecules with desired properties, such as binding affinity and selectivity to specific GPCRs. 7. Predicting GPCR properties: AI models can predict various properties of GPCRs, such as ligand binding sites (orthosteric, allosteric), activation mechanisms, and conformational changes. 8. Multi-target drug design: GPCRs are often involved in complex signaling networks. AI algorithms can integrate data from multiple sources, including genomics, proteomics, and pharmacological data, to identify potential drug targets within the GPCR signaling pathways. 9. Side effect prediction and clinical responses: AI can predict potential off-target effects and adverse drug reactions associated with GPCR-targeted drugs. With the revolution of biased signalling of GPCRs comes the possibility of designing drugs that selectively activate therapeutically important pathways over those that lead to undesired side effects. However, determination of ligand bias remains a major bottleneck requiring extensive experimental datasets and in vivo validation which does not always align with in vitro evidence (Kenakin, 2019). 10. Drug repurposing: AI algorithms can screen existing drugs and repurpose them for GPCR-related diseases. By analyzing drug-target interactions and disease pathways, AI can identify potential candidates that may have therapeutic effects on GPCRs. Pros and cons of current technology AI has the potential to revolutionize drug discovery by accelerating the process, optimizing drug design, and improving success rates. However, there are also certain cons associated with the use of AI in this field: 1) the quality and availability of data can be a challenge in AI-driven drug discovery; 2) the need for pattern recognition, often presented in the form of assumptions or hypotheses places similarity-based scoring functions at the core of any AI approach (Sanavia, Birolo, Montanucci, Turina, Capriotti & Fariselli, 2020) with intrinsic limitations (e.g. using sequence information alone to determine similarity cannot adequately predict protein structure (Jumper et al., 2021) ); 3) the same limitation holds when searching for novel receptor ligands, as “ligand-based approaches often bias molecule generation towards previously established chemical space, limiting their ability to identify truly novel chemotypes”(Thomas, Smith, O’Boyle, de Graaf & Bender, 2021) and 4) limit of input data can lead to a phenomenon called “the burden of sequence identity” (Sanavia, Birolo, Montanucci, Turina, Capriotti & Fariselli, 2020). The future According to the authors, the future of AI relies in 6 main areas: 1. Open-source data: accessibility of databases to a wider audience will encourage the development of new ideas and methods. 2. Greater application of unsupervised machine learning: most applications of ML in GPCR drug discovery have relied on traditional, hand-crafted feature construction, which limits the recognition of unknown patterns. Unsupervised deep learning that takes advantage of large unlabelled data (e.g., millions of compounds and assays on CHEMBL) to understand new proteins, ligands and their relationships is the future. 3. Interpretable machine learning: some AI models, such as deep learning neural networks, are often referred to as "black boxes" because they lack transparency and interpretability. Efforts will likely be made to develop AI models that can provide transparent explanations for their predictions and decisions. 4. Towards a comprehensive understanding of GPCRs, ligands, diseases, and their associations: open-access databases from experimental labs and new AI techniques will allow more associations and will promote multi-task learning. 5. Precision medicines for GPCRs: AI can facilitate the development of personalized treatments by analyzing patient data, including genomic information, clinical records, and treatment outcomes. By integrating this data with GPCR-related knowledge, AI can help identify patient-specific GPCR targets and optimize drug selection and dosing. 6. Automated tools for researchers: AI solutions which bring together theoretical, computational, and experimental labs will allow faster discovery and invention. While AI holds great potential, it is important to note that it is not a replacement for experimental validation and human expertise. Instead, it complements and assists researchers in the GPCR drug discovery process. Regulatory challenges, ethical considerations, and the need for interpretability of AI-driven models will need to be addressed to fully realize the potential of AI in GPCR drug discovery. The sharing and collaboration of data across academia, industry, and regulatory bodies will likely increase in the coming years and will facilitate the creation of larger and more diverse datasets for AI model training, improving their accuracy and generalizability. Check the original article at https://pubmed.ncbi.nlm.nih.gov/37161878/ #GPCR #DrGPCR #Ecosystem

Good day readers! We've got some GPCR updates to share with you. Take a look 👀! Save the date and join us next week, May 19th, 2023 for the Dr. GPCR Symposium on GPCR Activation and Signaling. If you want to present a poster, please submit your information using this form. Stay informed with the latest Dr. GPCR News by subscribing to our monthly newsletter. Below is your Classified GPCR News at a glance for May 1 to 7, 2023. GPCR Activation and Signaling Constitutive activity of the dopamine (D5 ) receptor, highly expressed in CA1 hippocampal neurons, selectively reduces CaV 3.2 and CaV 3.3 currents. Characterization of serotonin-5-HTR1E signaling pathways and its role in cell survival. Cholesterol Biases the Conformational Landscape of the Chemokine Receptor CCR3: A MAS SSNMR-Filtered Molecular Dynamics Study. Adrenomedullin 2/intermedin is a slow off-rate, long-acting endogenous agonist of the adrenomedullin2 G protein-coupled receptor. GPCR Binders, Drugs, and more Adenylyl cyclase 6 plays a minor role in the mouse inner ear and retina. GPCRs in Neuroscience GPCR interactions involving metabotropic glutamate receptors and their relevance to the pathophysiology and treatment of CNS disorders. G protein-coupled receptors in neurodegenerative diseases and psychiatric disorders. GPCRs in Oncology and Immunology Coronavirus Porcine Epidemic Diarrhea Virus Utilizes Chemokine Interleukin-8 to Facilitate Viral Replication by Regulating Ca2+ Flux. Methods & Updates in GPCR Research Multiplexed selectivity screening of anti-GPCR antibodies. Structural and Molecular Insights into GPCR Function Structural insight into an anti-BRIL Fab as a G-protein-coupled receptor crystallization chaperone. Molecular architecture of the Gαi-bound TRPC5 ion channel. Industry News Addex Therapeutics to Release Q1 2023 Financial Results and Host Conference Call on May 11, 2023 Neurocrine Biosciences Reports First Quarter 2023 Financial Results Find Therapeutics welcomes Dr. Jack Antel to its clinical advisory board Crinetics Pharmaceuticals First Quarter 2023 Earnings: Revenues Beat Expectations, EPS Lags “As Precision Medicine is critical, we cannot think about one size fits all” Dr Pina Cardarelli, Chief Scientific Officer, GPCR Therapeutics Trevena to Participate in Three Upcoming Conferences GPCR Events, Meetings, and Webinars ASPET 2023 - American Society for Pharmacology and Experimental Therapeutics (May 18 - 21, 2023). SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) PEGS Boston (May 15 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs Vice President, Medical Affairs Associate Scientist - Technology Senior Research Scientist/Principal Scientist- Technology Tissue Culture Scientist/Tissue Culture Manager Postdoctoral Researcher Director - Targeted Oncology Senior Scientist - Antibody Engineering Scientist—Immuno-Oncology Convergent Research - Senior Scientist, Cell-Based Assay Development Explore Dr. GPCR Ecosystem

Greetings fellow GPCR enthusiasts! This week's GPCR papers feature publications from Drs. Thomas P. Sakmar, Debbie L. Hay, Lukas Grätz, and more. Check out some of the latest GPCR discoveries below! Before you do so, mark your calendars and join us for the Dr. GPCR Symposium on GPCR Activation and Signaling on May 19th, 2023. If you're presenting a poster, submit your poster using the poster form. Also, keep yourself updated with the latest Dr. GPCR News by subscribing to our monthly newsletter. Below is your Classified GPCR News at a glance for April 24 to 30, 2023. Adhesion GPCRs Antiparallel dimer structure of CELSR cadherin in solution revealed by high-speed-atomic force microscopy. Tethered agonist activated ADGRF1 structure and signalling analysis reveal basis for G protein coupling. GPCR Activation and Signaling The ancestral ESCRT protein TOM1L2 selects ubiquitinated cargoes for retrieval from cilia. A visual opsin from jellyfish enables precise temporal control of G protein signalling. GPCR Binders, Drugs, and more Allosteric modulation of a human odorant receptor. Direct Selection of DNA-Encoded Libraries for Biased Agonists of GPCRs on Live Cells. Conformationally Selective 2-Aminotetralin Ligands Targeting the alpha2A- and alpha2C-Adrenergic Receptors. Bioorthogonal Tethering Enhances Drug Fragment Affinity for G Protein-Coupled Receptors in Live Cells. GPCRs in Cardiology, Endocrinology, and Taste Targeting G Protein-Coupled Receptors for Heart Failure Treatment. Single-cell transcriptome analysis of NEUROG3+ cells during pancreatic endocrine differentiation with small molecules. Novel roles for G protein-coupled receptor kinases in cardiac injury and repair. GPCRs in Neuroscience Evidence that RXFP4 is located in enterochromaffin cells and can regulate production and release of serotonin. GPCRs in Oncology and Immunology Stretch regulates alveologenesis and homeostasis via mesenchymal Gαq/11-mediated TGFβ2 activation. Single cell G-protein coupled receptor profiling of Transcription factor 21 expressing activated kidney fibroblasts. Small-molecule targeting of GPCR-independent noncanonical G-protein signaling in cancer. Methods & Updates in GPCR Research NanoBiT- and NanoBiT/BRET-based assays allow the analysis of binding kinetics of WNT-3A to endogenous Frizzled 7 in a colorectal cancer model. Reviews, GPCRs, and more G protein-coupled receptor-targeting antibody-drug conjugates: Current status and future directions. Genetic variants of G-protein coupled receptors associated with pubertal disorders. RGS proteins and their roles in cancer: friend or foe?. Structural and Molecular Insights into GPCR Function Cryo-EM structure of the endothelin-1-ETB-Gi complex. WNT stimulation induces dynamic conformational changes in the Frizzled-Dishevelled interaction. Industry News Orion Biotechnology will be participating in a panel discussion at the LSX World Congress Novel living yeast-based dual biosensor for detecting peptide variants The Galien Foundation Announces 2023 Prix Galien UK Award Candidates Exscientia Appoints Harvard Professor Franziska Michor, Ph.D. to Board of Directors GPCR Events, Meetings, and Webinars ASPET 2023 - American Society for Pharmacology and Experimental Therapeutics (May 18 - 21, 2023). SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) NEW PEGS Boston (May 15 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs NEW Associate Scientist - Technology NEW Senior Research Scientist/Principal Scientist- Technology NEW Tissue Culture Scientist/Tissue Culture Manager Postdoctoral Researcher Director - Targeted Oncology Senior Scientist - Antibody Engineering Scientist—Immuno-Oncology Convergent Research - Senior Scientist, Cell-Based Assay Development Explore Dr. GPCR Ecosystem

Hi friends!👋 Take a look at the latest and greatest in GPCR research. This week's GPCR papers feature authors such as Drs. Bryan Roth, Sudarshan Rajagopal, and Graeme Milligan. Save the date! Dr. GPCR Symposium on GPCR Activation and Signaling, May 19th, 2023. Submit your interest in giving a talk or presenting a poster to Hello@DrGPCR.com. Don't miss out! Just 3 weeks away! Stay updated with the latest Dr. GPCR News by subscribing to our monthly newsletter. Below is your Classified GPCR News at a glance for April 17 to 23, 2023. GPCR Activation and Signaling How can we improve the measurement of receptor signaling bias? Phosphorylation barcodes direct biased chemokine signaling at CXCR3. The relaxin receptor RXFP1 signals through a mechanism of autoinhibition. Subcellular location defines GPCR signal transduction. Regulation of the pro-inflammatory G protein-coupled receptor GPR84. GPCR Binders, Drugs, and more Development of a 5-HT7 receptor antibody for the rat: the good, the bad, and the ugly. GPCRs in Cardiology, Endocrinology, and Taste FFAR4 regulates cardiac oxylipin balance to promote inflammation resolution in HFpEF secondary to metabolic syndrome. GPCRs in Neuroscience G protein coupled receptors as targets for transformative neuropsychiatric therapeutics. GPCRs in Oncology and Immunology CCR6 as a Potential Target for Therapeutic Antibodies for the Treatment of Inflammatory Diseases. Methods & Updates in GPCR Research Solvent accessibility of a GPCR transmembrane domain probed by in-membrane chemical modification (IMCM). Structural and Molecular Insights into GPCR Function Lysophosphatidic acid, a simple phospholipid with myriad functions. Industry News Data Presented at AACR 2023 Highlights Exscientia’s Clinical and Preclinical Development The pathway of opioid drugs into cellular structures Inversago Pharma will present at BBHIC 2023 Orion Biotechnology attended the Swiss Biotech Day Addex Therapeutics attended the Swiss Biotech Day Sosei Heptares presented novel Computational Chemistry and Cheminformatics approaches for GPCR SBDD at UK QSAR Spring Meeting 2023 What the HEK? GPCR Events, Meetings, and Webinars ASPET 2023 - American Society for Pharmacology and Experimental Therapeutics (May 18 - 21, 2023). SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs NEW Postdoctoral Researcher Director - Targeted Oncology Senior Scientist - Antibody Engineering Scientist—Immuno-Oncology Convergent Research - Senior Scientist, Cell-Based Assay Development Post Doctoral Fellow Research associate in protein production Vice President, Oncology Clinical Research Explore Dr. GPCR Ecosystem

Transporting Protein-coupled receptors (GPCRs) to the synapse, where they are involved in neurotransmission, is a complex process involving several steps. From a general overview, after GPCRs synthesis in the endoplasmic reticulum, GPCRs are transported to the Golgi apparatus to undergo additional post-translational modifications and sorted for transport to their final destination. Several molecules from the SNARE complex, including vesicle-associated membrane protein 2 (VAMP2), regulate this last step toward the cell membrane [5,7]. VAMP2, also known as synaptobrevin, is a type of SNARE protein found on the synaptic vesicle membrane in neurons and is responsible for binding to the t-SNARE complex, a group of proteins found on the target membrane of the synapse. The t-SNARE complex is composed of two different proteins: syntaxin and SNAP-25; syntaxin is found on the target membrane of the synapse, while SNAP-25 is present on the membrane of the presynaptic neuron. The t-SNARE complex and VAMP2 interact to form the SNARE complex, which is essential for the fusion of the synaptic vesicle membrane with the target membrane, resulting in the release of neurotransmitters into the synapse. However, although the function of the SNARE protein complex in neurotransmitter release has been well characterized, the mechanisms that modulate the delivery of GPCRs to the membrane cell still need to be well known [3-5]. In the context of GPCR recycling, this process is complex and highly regulated that involves multiple molecular players, including SNARE proteins; here, SNARE proteins play a role in the fusion of recycling endosomes with the plasma membrane. Some studies have suggested that VAMP2 may be involved in regulating dopamine D2 receptor signaling by controlling the trafficking of the receptor to the cell surface. In addition, VAMP2 can interact with other GPCRs, such as the beta-2 adrenergic receptor and the mu-opioid receptor (MOR) [1,6,7]. However, the selectivity of SNARE complex proteins to regulate the release of different types of GPCRs during recycling is one of the questions still under investigation in the field. Through developing a high-resolution method, Hao Chen et al. directly visualized the fusion of vesicles containing GPCRs to the plasma membrane. With this technology, they evidenced the presence of VAMP2, specifically in recycling vesicles containing MOR but not B2AR or TFR in HEK293 cells and primary neurons. This study supports the idea that proteins from the fusion machinery are specific about the cargo molecules within the vesicles. These data are fascinating because in the case of MOR, which is a receptor with several splicing variants, its traffic to the membrane can be modified depending on the integrity of its bi-leucine sequence (which is considered a key element in its recycling), which can interact with different fusion proteins and also this different molecular codes will be modulated by different opioids, either endogenous or exogenous, promoving a differential organization of MOR receptors in vesicles with different proteins of the fusion machinery [6]. Since MOR receptor regulates pain perception and reward, the dysfunction in the MOR-SNARE complex interaction can lead to various neurological disorders, such as chronic pain and addiction. Therefore, their comprehension is essential for developing new treatments for these disorders and advancing our understanding of the brain and nervous system. You can consult the article at the following link: https://www.ecosystem.drgpcr.com/structural-and-molecular-insights-into-gpcr-function/vesicle-associated-membrane-protein-2-is-a-cargo-selective-v-snare-for-a-subset-of-gpcrs #GPCR #DrGPCR#Ecosystem Crilly, S.E., W. Ko, Z.Y. Weinberg, and M.A. Puthenveedu. 2021. Conformational specificity of opioid receptors is determined by subcellular location irrespective of agonist. Elife. 10:e67478. Drake, M.T., S.K. Shenoy, and R.J. Lefkowitz. 2006. Trafficking of G protein coupled receptors. Circ. Res. 99:570–582. Jurado, S., D. Goswami, Y. Zhang, A.J.M. Molina, T.C. Südhof, and R.C. Malenka. 2013. LTP requires a unique postsynaptic SNARE fusion machinery. Neuron. 77:542–558. Schoch, S., F. Deak, A. K ´ onigstorfer, M. Mozhayeva, Y. Sara, T.C. Südhof, and ¨ E.T. Kavalali. 2001. SNARE function analyzed in synaptobrevin/VAMP knockout mice. Science. 294:1117–1122. 5. Neurotransmitter release: The last millisecond in the life of a synaptic vesicle. Neuron. 80:675–690. Wang, F., X. Chen, X. Zhang, and L. Ma. 2008. Phosphorylation state of muopioid receptor determines the alternative recycling of receptor via Rab4 or Rab11 pathway. Mol. Endocrinol. 22:1881–1892. Wickner, W., and R. Schekman. 2008. Membrane fusion. Nat. Struct. Mol. Biol. 15:658–664.

Hey there! Are you ready for some exciting news in the GPCR world? Then check this out 👓! Now you can submit your job opening and offer a position in an exclusive GPCR community around the globe with ease by using this form. Save the date! Join us for the upcoming Dr. GPCR Symposium on GPCR Activation and Signaling on May 19th, 2023. While the program is being finalized, if you're interested in giving a talk or presenting a poster, please email us at Hello@DrGPCR.com. Don't miss out on this exciting opportunity! Subscribe to our monthly newsletter for more Dr. GPCR News. Below is your Classified GPCR News at a glance for April 10 to 16, 2023. Adhesion GPCRs Downstream signaling of the disease associated mutations on GPR56/ADGRG1. GPCR Activation and Signaling Ligand-dependent intracellular trafficking of the G protein-coupled P2Y6 receptor. GPCR Binders, Drugs, and more Discovery of SYD5115, a novel orally active small molecule TSH-R antagonist. GPCRs in Cardiology, Endocrinology, and Taste G-protein coupled receptor 19 (GPR19) knockout mice display sex-dependent metabolic dysfunction. GPCRs in Neuroscience GPR160 is not a receptor of anorexigenic cocaine- and amphetamine-regulated transcript peptide. Neurotensin receptor 1-biased ligand attenuates neurotensin-mediated excitation of ventral tegmental area dopamine neurons and dopamine release in the nucleus accumbens. Multiple subthreshold GPCR signals combined by the G proteins Gαq and Gαs activate the C. elegans egg-laying muscles. GPCRs in Oncology and Immunology G Protein-coupled Receptor-mediated Membrane Targeting of PLCγ2 is Essential for Neutrophil Chemotaxis. Methods & Updates in GPCR Research Activity Map and Transition Pathways of G Protein-Coupled Receptor Revealed by Machine Learning. Computational insights into ligand-induced G protein and β-arrestin signaling of the dopamine D1 receptor. In situ visualization of opioid and cannabinoid drug effects using phosphosite-specific GPCR antibodies. Reviews, GPCRs, and more Internal and external modulation factors of the orexin system (REVIEW). Structural and Molecular Insights into GPCR Function Function and dynamics of the intrinsically disordered carboxyl terminus of β2 adrenergic receptor. Transmembrane protein CD69 acts as an S1PR1 agonist. Industry News A Current View of Allosteric GPCR Drug Discovery from the 2nd Annual GPCRs-Targeted Drug Discovery Summit, Boston MA Pathios Therapeutics Unveils PTT-4256, a Highly Potent and Selective Inhibitor of GPR65, in Presentation at American Association for Cancer Research (AACR) Annual Meeting 2023 Setting GPCRs free Dr. Michel Bouvier received the title of Doctor Honoris Causa from the Université de Montpellier Domain Therapeutics presented 3 posters at AACR 2023 Exscientia presented a poster at AACR 2023 Pathios Therapeutics presented a poster at AACR 2023 GPCR Events, Meetings, and Webinars NEW FREE 7th ERNEST GPCR zoominar (April 20, 2023) SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) Swiss Biotech Day (April 24 - 25, 2023) ASPET 2023 - American Society for Pharmacology and Experimental Therapeutics (May 18 - 21, 2023). SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs NEW Director - Targeted Oncology NEW Senior Scientist - Antibody Engineering NEW Scientist—Immuno-Oncology Convergent Research - Senior Scientist, Cell-Based Assay Development Post Doctoral Fellow Research associate in protein production Vice President, Oncology Clinical Research PostDoctoral Associate PostDoctoral Position Team Lead protein production and profiling Explore Dr. GPCR Ecosystem

Hello, fellow GPCR enthusiasts! 👋 Check out the latest advancements in GPCR studies. For Dr. GPCR News, please subscribe to our monthly newsletter. Also, please mark your calendar for the next Dr. GPCR Symposium on GPCR Activation and Signaling held on May 19th, 2023. The program is under construction but if you’d like to give a talk or present a poster, email us at Hello@DrGPCR.com Below is your Classified GPCR News at a glance for April 3rd to 9th, 2023. GPCR Activation and Signaling Phosphorylation barcodes direct biased chemokine signaling at CXCR3. SUCNR1 signaling in adipocytes controls energy metabolism by modulating circadian clock and leptin expression. GABABR silencing of nerve terminals. GPCR Binders, Drugs, and more Inhibiting a promiscuous GPCR: iterative discovery of bitter taste receptor ligands. The molecular basis of the antidepressant action of the magic mushroom extract, psilocin. Sodium is a negative allosteric regulator of the ghrelin receptor. GPCRs in Cardiology, Endocrinology, and Taste Novel roles for G protein-coupled receptor kinases in cardiac injury and repair. DRD1 is exempt from TGFβ-mediated antifibrotic GPCR landscape tampering in lung fibroblasts. βHB inhibits glucose-induced GLP-1 secretion in GLUTag and human jejunal enteroids. GPCRs in Neuroscience G protein-coupled receptors (GPCRs) as Potential Therapeutics for Psychiatric Disorders. Local 5-HT signaling bi-directionally regulates the coincidence time window for associative learning. GPCR-mediated calcium and cAMP signaling determines psychosocial stress susceptibility and resiliency. GPR37L1 controls maturation and organization of cortical astrocytes during development. Methods & Updates in GPCR Research The Road to Quantitative Lipid Biochemistry in Living Cells. Neuronal activity-induced, equilibrative nucleoside transporter-dependent, somatodendritic adenosine release revealed by a GRAB sensor. Reviews, GPCRs, and more Heterotrimeric G proteins regulate planarian regeneration and behavior. Transcriptomic profiling of sex-specific olfactory neurons reveals subset-specific receptor expression in Caenorhabditis elegans. β- Arrestins: Structure, Function, Physiology, and Pharmacological Perspectives. Structural and Molecular Insights into GPCR Function Sub-millisecond conformational dynamics of the A2A adenosine receptor revealed by single-molecule FRET. Vesicle-associated membrane protein 2 is a cargo-selective v-SNARE for a subset of GPCRs. Industry News Orion Biotechnology at the AACR 2023. Eurofins Discovery Acquires CALIXAR, Expanding Integrated Drug Discovery Capabilities and Expertise. Domain Therapeutics to present three posters at AACR 2023 annual meeting. GPCR Events, Meetings, and Webinars NEW AACR 2023 - American Association for Cancer Research (April 14 - 19, 2023). SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) Swiss Biotech Day (April 24 - 25, 2023) NEW ASPET 2023 - American Society for Pharmacology and Experimental Therapeutics (May 18 - 21, 2023). SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs Convergent Research - Senior Scientist, Cell-Based Assay Development Post Doctoral Fellow Research associate in protein production Vice President, Oncology Clinical Research PostDoctoral Associate PostDoctoral Position Team Lead protein production and profiling Pharmacologist Postdoctoral positions at UC San Diego Medical Director CADD and Informatics Head or Team Lead (depending on experience) with Structural Bioinformatics Expertise Explore Dr. GPCR Ecosystem

Canonical chemokine receptors – scavenging “decoys” Chemokine receptors coordinate cell migration upon interaction with their cognate chemokines. Upon activation, chemokine receptors coupe to the Gαi class of heterotrimeric G proteins, which, in turn, activate signaling pathways that ultimately lead to cell migration. The chemokine receptor family comprises 19 canonical chemokine receptors together with 4 atypical chemokine receptors (ACKRs) which do not couple to G proteins and behave as scavenging “decoys” in order to either limit chemokines spatial availability or to remove them from in vivo sites, while maintaining the responsiveness of canonical G protein–coupled chemokine receptors that bind to the same ligand(s) (R. J. B. Nibbs et al. 2013). Although less characterized, canonical chemokine receptor systems have been also shown to not only directly regulatE migration but also play a scavenging role (e.g. CCR2, CXCR2, CXCR3, and CX3CR1) (A. E. Cardona et al. 2008). CCR2 is an example of a dual-function receptor that directly regulates both cell migration and scavenging (S. Volpe et al. 2012). Molecular signature of CCR2 scavenging - no G proteins, no GRKs, no arrestins, and no clathrin Chemokine scavenging mechanism involves internalization and recycling of the receptor with clearance of the ligand from the extracellular space however, the molecular cues involved in this process are poorly characterized. In this study, the molecular signature of CCR2 scavenging role is investigated. CCR2 canonical signaling requires the activation of Gαiβγ, followed by phosphorylation of the receptor C terminus by G protein receptor kinases (GRKs), specifically GRK2 and GRK3 (A. M. Aragay et al. 1998). Then β-Arrestin recruitment takes place allowing receptor internalization through clathrin-mediated pits (V. V. Gurevich, et al. 2018; Y. A. Berchiche et al. 2011). In this study, the role of these 3 molecular players (G proteins, GRKs, and β-arrestin) is investigated. Removal of G proteins by using CRISPR KO of Gαi (Gαi KO) or KO of all Gα subtypes (Gα_all KO) (M. Grundmann et al. 2018), did not affect chemokine scavenging, consistent with prior work (S. Volpe et al. 2012; B. N. Zhao et al. 2019). Recruitment of both β-arrestin1 and β-arrestin2 was significantly diminished iin Gαi KO and Gα_all KO HEK293 cells, however there was only a minor loss in receptor internalization, which is surprising since internalization is known to be a consequence of β-arrestin recruitment. In addition, constitutive internalization of CCR2 was shown to be G protein-independent by a “prelabel” flow cytometry assay (B. N. Zhao et al. 2019) together with confocal fluorescence microscopy. Regarding GRKs role in CCR2 scavenging, BRET, flow cytometry and fluorescence microscopy readouts revealed that CCL2-dependent internalization was partially decreased in GRK2/3 KO cells and almost completely lost in the GRK2/3/5/6 KO cells, whereas constitutive receptor internalization was unaffected. Moreover, HEK293 cells with CRISPR KO of both β-arrestin1 and β-arrestin2 only led to a small but measurable loss of CCL2 scavenging and did not affect constitutive receptor internalization, with the same observations made for CCR2-expressing THP-1 monocytic cell line, together with an increase in migration, consistent with the role of β-arrestin in receptor desensitization. Although some GPCRs constitutively internalize through clathrin-coated pits independently of phosphorylation and β-arrestin (M. M. Paing et al. 2022; J. L. Parent, et al. 2001), inhibition of clathrin-mediated endocytosis (CME) did not affect the ability of CCR2 to scavenge chemokine and had only a small effect on constitutive internalization. Using fast recycling, slow recycling, and late endosomal recycling markers, CCR2 was also shown to be rapidly sorted into fast-recycling Rab4-positive endosomes in a more sustained manner compared to the atypical scavenging receptor ACKR3, and Rab7-positive late endosomes. CCR2 was also shown to be resistant to degradation and most likely recycling through the trans-Golgi network (TGN) where Rab7 is associated (A. Sapmaz et al. 2019, M. N. J. Seaman 2012). The ability to rapidly recycle, which was shown by a chemokine washout BRET assay (Y. Namkung et al. 2016), and avoid depletion may also contribute to its ability to efficiently scavenge chemokine. CCR2 vs CCR1 scavenging signatures CCR1 is constitutively phosphorylated, constitutively interacts with β-arrestin2, and constitutively internalizes in a β-arrestin2–dependent manner (C. T. Gilliland et. al 2013). In line, β-Arrestin KO cells showed a major reduction in CCL14 scavenging by CCR1, in contrast with a minor effect observed for CCR2. β-arrestin KO cells revealed the rapid reassociation of Gαi and Gβγ on CCR1, in contrast with CCR2, indicating that β-arrestin regulates scavenging and signaling of CCR1 to a greater extent than CCR2. Functional relevance and implications of scavenging of canonical chemokine receptors Clearing circulating chemokines and chemokines from tissue microenvironments has been well described for ACKRs which work in a team with canonical chemokine receptor to drive regulated immunological responses and inflammatory conditions, preventing overstimulation and activation. However, this scavenging role has been also described for several canonical chemokine receptors, including CCR2. In this study, it is revealed that CCR2 scavenging role was largely independent of the classical GPCR signaling pathways, revealing the existence of two distinct functional populations – one that directs cell migration and one that regulates scavenging. But why do canonical chemokine receptors have this dual role in physiology? Scavenging may allow cells to continuously migrate by remaining responsive to chemokines (S. Volpe et al. 2012); it dampens the inflammatory response when needed (C. A. H. Hansell et al. 2011); and it may interfere with other chemokine receptors which share the ligands and affect cell migration (A. E. Cardona et al. 2008). However, it is important to note that this scavenging role does not occur in all chemokine receptor systems such as CXCR4, which relies on ACKRs. How balanced is this dual role? In monocytes and dendritic cells exposed to treatments mimicking inflammation, CCR1, CCR2, and CCR5 switch purely to scavenging (G. D’Amico et. al 2000), becoming incapable of promoting cell migration, a phenomenon which is likely to be mediated by changes in the cell motility machinery with receptor-specific switches not yet described to play a role. What are the implications in drug discovery? This scavenging function should be considered when evaluating the safety and therapeutic efficacy of blocking receptor-ligand binding. CCR2 inhibition leads to inhibition of scavenging and elevated plasma levels of CCL2 (Y. Wang et al. 2009; R. J. Aiello et. al 2010) which may ultimately compete with receptor antagonists, thereby decreasing the efficacy (J. Gilbert et. al 2011). This study provides a comprehensive analysis of CCR2 scavenging role however to fully understand the role of canonical chemokine receptors scavenging population pathophysiology, a better understanding of the regulatory mechanisms will be required. Check the original article at https://pubmed.ncbi.nlm.nih.gov/36719944/ #GPCR #DrGPCR#Ecosystem

Hello Readers👋, This week in the GPCR news we have collected the most up-to-date news for you. Enjoy! For more great content, subscribe to our Dr. GPCR News monthly newsletter. Also, please mark your calendar for the next Dr. GPCR Symposium on GPCR Activation and Signaling held on May 19th, 2023. The program is under construction but if you’d like to give a talk or present a poster, email us at Hello@DrGPCR.com Below is your Classified GPCR News at a glance for March 27th to April 4th, 2023. Adhesion GPCRs Optimized genetic code expansion technology for time-dependent induction of adhesion GPCR-ligand engagement. The adhesion G-protein-coupled receptor VLGR1/ADGRV1 controls autophagy. GPCR Activation and Signaling Butyrate potentiates Enterococcus faecalis lipoteichoic acid-induced inflammasome activation via histone deacetylase inhibition. Negative allosteric modulation of the glucagon receptor by RAMP2. Itch receptor MRGPRX4 interacts with the receptor activity-modifying proteins (RAMPs). Regulator of G-Protein Signalling 4 (RGS4) negatively modulates nociceptin/orphanin FQ opioid receptor signalling: Implication for l-Dopa-induced dyskinesia. Cannabinoid Tolerance in S426A/S430A x β-Arrestin 2 Knockout Double-Mutant Mice. Gαi-derived peptide binds the µ-opioid receptor. Deficiency of G protein-coupled receptor Gpr111/Adgrf2 causes enamel hypomineralization in mice by alteration of the expression of kallikrein-related peptidase 4 (Klk4) during pH cycling process. GPCR Binders, Drugs, and more Predicting allosteric sites using fast conformational sampling as guided by coarse-grained normal modes. Statins inhibit protein kinase D (PKD) activation in intestinal cells and prevent PKD1-induced growth of murine enteroids. GPCRs in Cardiology, Endocrinology, and Taste Pepducin ICL1-9-Mediated β2-Adrenergic Receptor-Dependent Cardiomyocyte Contractility Occurs in a Gi Protein/ROCK/PKD-Sensitive Manner. Dietary compounds activate an insect gustatory receptor on enteroendocrine cells to elicit myosuppressin secretion. GPCRs in Neuroscience Arrestin-dependent nuclear export of phosphodiesterase 4D promotes GPCR-induced nuclear cAMP signaling required for learning and memory. Neuronal activity-induced, equilibrative nucleoside transporter-dependent, somatodendritic adenosine release revealed by a GRAB sensor. GPCRs in Oncology and Immunology Receptor autoantibodies: Associations with cardiac markers, histology, and function in human non-ischaemic heart failure. Methods & Updates in GPCR Research Discovery and design of G protein-coupled receptor targeting antibodies. A Vaccinia-based system for directed evolution of GPCRs in mammalian cells. First complete genome sequence of lumpy skin disease virus directly from a clinical sample in South India. Reviews, GPCRs, and more NMR applications to GPCR recognition by peptide ligands. Developmental and homeostatic signaling transmitted by the G-protein coupled receptor FPR2. Dopamine Receptor D1R and D3R and GRK4 Interaction in Hypertension. Therapeutic potential of opioid receptor heteromers in chronic pain and associated comorbidities. Investigating the potential of GalR2 as a drug target for neuropathic pain. The impact of cryo-EM on determining allosteric modulator-bound structures of G protein-coupled receptors. Structural and Molecular Insights into GPCR Function Predicted Three-Dimensional Structure of the GCR1 Putative GPCR in Arabidopsis thaliana and Its Binding to Abscisic Acid and Gibberellin A1. Structural insights into constitutive activity of 5-HT6 receptor. Industry News Confo Therapeutics Enters into Collaboration Agreement with Daiichi Sankyo to Discover Novel Medicines for CNS Diseases Addex Reports Full Year 2022 Financial Results and Provides Corporate Update Sosei Heptares Announces the Publication of its Inaugural ESG Report, for the Financial Year ended 31 December 2022 Addex Raises $5.0 Million in Equity Financing Testing the limits of SMILES-based de novo molecular generation with curriculum and deep reinforcement learning Structure Therapeutics Reports Fourth Quarter and Full Year 2022 Financial Results and Recent Highlights Salipro Biotech publishes research in Scientific Reports on structure-function studies for membrane proteins in collaboration with AstraZeneca Expansion of precision oncology pipeline announced by Exscientia Upcoming start-up ecosystem - leadXpro and InterAx part of Fujitsu CaaS GPCR Events, Meetings, and Webinars SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) Swiss Biotech Day (April 24 - 25, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs NEW Convergent Research - Senior Scientist, Cell-Based Assay Development NEW Post Doctoral Fellow NEW Research associate in protein production NEW Vice President, Oncology Clinical Research NEW PostDoctoral Associate NEW PostDoctoral Position Team Lead protein production and profiling Pharmacologist Postdoctoral positions at UC San Diego Medical Director CADD and Informatics Head or Team Lead (depending on experience) with Structural Bioinformatics Expertise Explore Dr. GPCR Ecosystem

Hello Readers👋, We're excited to have you in the Dr. GPCR Ecosystem. Every week, we send you the most relevant news, research, and advancements in the field. For Dr. GPCR News, subscribe to our monthly newsletter. This month on the Dr. GPCR News video edition, we chatted with Dr. Oliver Hartley. Below is your Classified GPCR News at a glance for March 20th to 26th, 2023. GPCR Activation and Signaling Targeting biased signaling by PAR1: Function and molecular mechanism of parmodulins. The Wnt pathway protein Dvl1 targets Somatostatin receptor 2 for lysosome-dependent degradation. Loss of biased signaling at a G protein-coupled receptor in overexpressed systems. GPCR Binders, Drugs, and more A snake toxin as a theranostic agent for the type 2 vasopressin receptor Small-Molecule Fluorescent Ligands for the CXCR4 Chemokine Receptor. GPCRs in Cardiology, Endocrinology, and Taste Green mamba peptide targets type-2 vasopressin receptor against polycystic kidney disease GPCRs in Oncology and Immunology Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion. CCL5-producing migratory dendritic cells guide CCR5+ monocytes into the draining lymph nodes. Methods & Updates in GPCR Research The application of targeted protein degradation technologies to G protein-coupled receptors. Structural and Molecular Insights into GPCR Function Physiological changes in bilayer thickness induced by cholesterol control GPCR rhodopsin function. Evidence that RXFP4 is located in enterochromaffin cells and can regulate production and release of serotonin. Ultrafast structural changes direct the first molecular events of vision. New insights into GPCR coupling and dimerisation from cryo-EM structures. Industry News Domain Therapeutics was one of the 4 Biotech Trophy nominees at 2023 HealthTech awards. The delisting dilemma: Why do so many biotechs face being kicked off the Nasdaq?. Exscientia Business and Financial Update for the Full Year 2022. Sosei Heptares Confirms Changes to its Board and Executive Management Team at Annual Shareholders’ Meeting. Function Therapeutics has been awarded a Phase 1 SBIR grant from NSF. Function Therapeutics has a new website. Call for GPCR Papers GPCRs: Signal Transduction. Ends tomorrow - March 31st, 2023. GPCR Events, Meetings, and Webinars FREE 5th ERNEST GPCR ECI zoominar. (March 30, 2023) NEW Webinar - Antiverse: Engineering the Future of Drug Discovery (March 30, 2023) NEW Molecular Dynamics in Pharma (March 31, 2023) NEW FREE Workshop - Challenges in GPCR Drug Discovery (March 31, 2023) SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) Swiss Biotech Day (April 24 - 25, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). NEW FREE Symposium - IPI Surfacing (June 15, 2023) Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) FREE 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs NEW Team Lead protein production and profiling NEW Pharmacologist Postdoctoral positions at UC San Diego Medical Director CADD and Informatics Head or Team Lead (depending on experience) with Structural Bioinformatics Expertise Scientist Corporate Strategy Analyst PhD candidate or Postdoc. Director, Head of PMO. Associate Scientist, Protein Science. Explore Dr. GPCR Ecosystem

Hello Readers👋, Welcome to our GPCR weekly newsletter! We thrive to keep you up-to-date with everything happening in the world of GPCR. Remember, you can always adjust your email preferences in your account settings. For Dr. GPCR News, please subscribe to our monthly newsletter. The first Dr. GPCR Symposium starts tomorrow! If you'd like to present a poster, please submit yours today. There is still time. There is no limit to the number of posters we can accommodate, so hurry and submit your poster so that we can add it to the list of posters and increase the number of people who can ''stop by'' it tomorrow. Below is your Classified GPCR News at a glance for March 13th to 19th, 2023. Adhesion GPCRs Crystal Structure of the Extracellular Domains of GPR110. Monitoring paxillin in astrocytes reveals the significance of the adhesion GPCR VLGR1/ADGRV1 for focal adhesion assembly. GPCR Activation and Signaling Components of TOR and MAP kinase signaling control chemotropism and pathogenicity in the fungal pathogen Verticillium dahliae. A Rab10-ACAP1-Arf6 GTPases cascade modulates M4 muscarinic acetylcholine receptor trafficking and signaling. Neurotensin Receptor Allosterism Revealed in Complex with a Biased Allosteric Modulator. α1-adrenoceptor ligands inhibit chemokine receptor heteromerization partners of α1B/D-adrenoceptors via interference with heteromer formation. The immunometabolite itaconate stimulates OXGR1 to promote mucociliary clearance during the pulmonary innate immune response. GPCR Binders, Drugs, and more Design of Drug Efficacy Guided by Free Energy Simulations of the β2-Adrenoceptor. Pharmacological characterization of novel small molecule agonists and antagonists for the orphan receptor GPR139. A small molecule ligand for the novel pain target, GPR171, produces minimal reward in mice. Monitoring the Reversibility of GPCR Signaling by Combining Photochromic Ligands with Label-free Impedance Analysis. GPCRs in Cardiology, Endocrinology, and Taste Structure-based design of novel melanin-concentrating hormone receptor-1 ligands based on saturated nitrogen-containing heterocycles. GPCRs in Neuroscience Physiological Condition-Dependent Changes in Ciliary GPCR Localization in the Brain. Endogenous l- to d-amino acid residue isomerization modulates selectivity between distinct neuropeptide receptor family members. GPCRs in Oncology and Immunology Expanding role of CXCR2 and therapeutic potential of CXCR2 antagonists in inflammatory diseases and cancers. Reviews, GPCRs, and more The leukotriene B4 receptors BLT1 and BLT2 as potential therapeutic targets. New paradigms in purinergic receptor ligand discovery. G protein-coupled receptor pharmacology - insights from mass spectrometry. Structural and Molecular Insights into GPCR Function Structural basis of selective cannabinoid CB2 receptor activation. Structural basis for motilin and erythromycin recognition by motilin receptor. Industry News Addex Announces Upcoming Conferences It Will Participate For H1 2023 Learn more about Exscientia new pipeline candidate, EXS74539 Domain Therapeutics unveiled its 2023 women-men professional equality index Octant introduces Hypatia, their New Robotic Drug Hunter GPCR Therapeutics Announces Publication in Nature Scientific Reports on Novel Anticancer Therapy An extraordinary opportunity to generate proprietary chemo-informatics for GPCR drug discovery - Design Pharmaceuticals Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023. GPCR Events, Meetings, and Webinars NEW 4th ERNEST GPCR ECI zoominar. (March 23, 2023) NEW 5th ERNEST GPCR ECI zoominar. (March 30, 2023) SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) NEW Swiss Biotech Day (April 24 - 25, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). NEW Training School on “Cell-based assays to study Adhesion GPCR function". (June 28 - 30, 2023) 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs Postdoctoral positions at UC San Diego Medical Director CADD and Informatics Head or Team Lead (depending on experience) with Structural Bioinformatics Expertise Scientist Corporate Strategy Analyst PhD candidate or Postdoc. Director, Head of PMO. Associate Scientist, Protein Science. Explore Dr. GPCR Ecosystem

Hello Readers👋, Thank you for being here with us this week. We work tirelessly to bring you informative and exciting news about the world of GPCR every Thursday. To get the latest, Dr. GPCR News, don't forget to sign up for our monthly newsletter. Are you ready for our first Dr. GPCR Symposium event held on March 24th? If you'd like to present a poster, please submit yours today. There is no limit to the number of posters we can accommodate, so hurry and submit your poster so that we can add it to the list of posters and increase the number of people who can ''stop by'' it on March 24th, 2023. Below is your Classified GPCR News at a glance for March 6th to 12th, 2023. GPCR Activation and Signaling Autoregulation of GPCR signalling through the third intracellular loop. GPCR Binders, Drugs, and more Autoantibodies targeting G protein-coupled receptors: An evolving history in autoimmunity. Report of the 4th international symposium. Discovery, synthesis and mechanism study of 2,3,5-substituted [1,2,4]-thiadiazoles as covalent inhibitors targeting 3C-Like protease of SARS-CoV-2. Novel Xanomeline-Containing Bitopic Ligands of Muscarinic Acetylcholine Receptors: Design, Synthesis and FRET Investigation. GPCRs in Neuroscience Quinpirole ameliorates nigral dopaminergic neuron damage in Parkinson's disease mouse model through activating GHS-R1a/D2R heterodimers. The crosstalk between 5-HT2AR and mGluR2 in schizophrenia. GPCRs in Oncology and Immunology GPR176 Promotes Cancer Progression by Interacting with G Protein GNAS to Restrain Cell Mitophagy in Colorectal Cancer. Intermediate-state-trapped mutants pinpoint G protein-coupled receptor conformational allostery. Structural and Molecular Insights into GPCR Function Endogenous l- to d-amino acid residue isomerization modulates selectivity between distinct neuropeptide receptor family members. Molecular sensing of mechano- and ligand-dependent adhesion GPCR dissociation. Structural basis of peptide recognition and activation of endothelin receptors. Industry News Professor Dame Carol Robinson Received 2023 ASMS John.B.Fenn Award Exscientia Announces Expansion of its Precision Oncology Pipeline Exscientia to Present Data Highlighting Pipeline and Precision Medicine Platform at AACR Fast Company has named Exscientia as one of the Most Innovative Companies of 2023 Dirk Loeffert CEO & Founder DesignPharma Interview GPCR-targeting drugs: A renewed focus on a ubiquitous group of proteins arcoscreen will be at Bio-Europe 2023 in Basel. Domain Therapeutics was nominated for the Trophée de l'Innovation Technologique (Technological Innovation Award) Sosei Heptares Receives Approval for Change of Market Listing Segment to the Tokyo Stock Exchange Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023. GPCR Events, Meetings, and Webinars GEM2023. (March 14-17, 2023). SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs CADD and Informatics Head or Team Lead (depending on experience) with Structural Bioinformatics Expertise Scientist Corporate Strategy Analyst PhD candidate or Postdoc. Director, Head of PMO. Associate Scientist, Protein Science. PhD Studentship - Integrated Approaches In Pharmacology, Computer Simulations And Machine Learning To Predict Ligand Signaling Pathways Via The Anti-Inflammatory Receptor Gpr84. PhD Studentship - Insight Into The Neuroprotective Function Of The Prostaglandin Receptor Ep2. Explore Dr. GPCR Ecosystem

Hello Readers👋, GPCR weekly newsletter is here! Prepare to stay informed of the most recent findings, developments, and news in the industry. Keep in mind that you can always change your email preferences in your account settings. Join us in our monthly newsletter to receive Dr. GPCR News. Great News! We are excited to welcome you to the first Dr. GPCR Symposium event held on March 24th. If you'd like to present a poster, please submit yours today. There is no limit to the number of posters we can accommodate, so hurry and submit your poster so that we can add it to the list of posters and increase the number of people who can ''stop by'' it on March 24th, 2023. Below is your Classified GPCR News at a glance for February 27th to March 5th, 2023. GPCRs in Cardiology, Endocrinology, and Taste Dietary compounds activate an insect gustatory receptor on enteroendocrine cells to elicit myosuppressin secretion. GPCRs in Neuroscience Physiological Condition Dependent Changes in Ciliary GPCR Localization in the Brain. Establishment of a CaCC-based Cell Model and Method for High-throughput Screening of M3 Receptor Drugs. Publisher Correction: Transcriptional adaptation of olfactory sensory neurons to GPCR identity and activity. Elucidation of a dynamic interplay between a beta-2 adrenergic receptor, its agonist, and stimulatory G protein. GPCRs in Oncology and Immunology GPR143 controls ESCRT-dependent exosome biogenesis and promotes cancer metastasis. Methods & Updates in GPCR Research Time- and cost-efficient bacterial expression and purification of potato apyrase. FSHR activation through small molecule modulators: Mechanistic insights from MD simulations. Reviews, GPCRs, and more Short-chain fatty acids: possible regulators of insulin secretion. Function and structure of bradykinin receptor 2 for drug discovery. Developing novel antifungals: lessons from G protein-coupled receptors. Function and regulation of GPR84 in human neutrophils. Structural and Molecular Insights into GPCR Function The activation mechanism and antibody binding mode for orphan GPR20. Structural details of a Class B GPCR-arrestin complex revealed by genetically encoded crosslinkers in living cells. Industry News Design Pharmaceuticals has a new website Confo Therapeutics Announces Global Licensing Agreement with Lilly for Peripheral Pain Candidate, CFTX-1554 Cardiff-based Antiverse raises €2.8M for its computational antibody drug discovery platform Antiverse identifies therapeutic antibodies targeting GPCRs CEO Andrew Hopkins on Exscientias AI-Driven Approach to Modern Drug Discovery Arcoscreen will present in the start-up village at Future Labs Live Basel Exscientia Announces Collaboration with Charité to Advance Development of Precision Medicine Platform Sosei Heptares and Neurocrine Biosciences won Top Out-Licensing Deal on the 8th annual Japan Deal of the Year Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023. GPCR Events, Meetings, and Webinars ERNEST ECI zoominar. (March 9) GEM2023. (March 14-17, 2023). SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 11th Adrenoceptor Symposium: Adrenoceptors and GPCR Signalling (June 30 - July 1, 2023) 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). 3rd Annual Meeting IRN I-GPCRNet (October 25 - 27, 2023) GPCR Jobs PhD candidate or Postdoc. Director, Head of PMO. Associate Scientist, Protein Science. PhD Studentship - Integrated Approaches In Pharmacology, Computer Simulations And Machine Learning To Predict Ligand Signaling Pathways Via The Anti-Inflammatory Receptor Gpr84. PhD Studentship - Insight Into The Neuroprotective Function Of The Prostaglandin Receptor Ep2. Computational drug design PhD student at the Department of Drug Design and Pharmacology. Computational drug design postdoc at the Department of Drug Design and Pharmacology. Software / database development PhD student at Department of Drug Design and Pharmacology. Software / database development postdoc at the Department of Drug Design and Pharmacology. Explore Dr. GPCR Ecosystem

Hello Readers👋, Thank you for joining us this week. Each week on Thursday, you receive informative and exciting news about GPCRs. And don't forget, for Dr. GPCR news content, make sure to subscribe to our monthly newsletter for Dr. GPCR News. Sometimes we may miss publications. If you'd like to suggest papers to include in our news, please email them to us at Hello@DrGPCR.com. Below is your Classified GPCR News at a glance for February 20th to 26th, 2023. GPCR Activation and Signaling Coupling between GPR143 and dopamine D2 receptor is required for selective potentiation of dopamine D2 receptor function by L-3,4-dihydroxyphenylalanine in the dorsal striatum. All-Atom Molecular Dynamics Simulations Indicated the Involvement of a Conserved Polar Signaling Channel in the Activation Mechanism of the Type I Cannabinoid Receptor. Allosteric modulation of conserved motifs and helices in 5HT2BR: Advances drug discovery and therapeutic approach towards drug resistant epilepsy. Molecular basis for differential activation of p101 and p84 complexes of PI3Kγ by Ras and GPCRs. Impact of membrane lipid polyunsaturation on dopamine D2 receptor ligand binding and signaling GPCR Binders, Drugs, and more Discovery of the Potent and Selective MC4R Antagonist PF-07258669 for the Potential Treatment of Appetite Loss. Small-molecule targeting of GPCR-independent non-canonical G protein signaling inhibits cancer progression. Application of computational methods for class A GPCR Ligand discovery. Gαi-derived peptide binds the µ-opioid receptor. GPCRs in Cardiology, Endocrinology, and Taste α1-Adrenergic Receptors: Insights into Potential Therapeutic Opportunities for COVID-19, Heart Failure, and Alzheimer's Disease. Gβγ subunits co-localize with RNA polymerase II and regulate transcription in cardiac fibroblasts. Slow-rising and fast-falling dopaminergic dynamics jointly adjust negative prediction error in the ventral striatum. Methods & Updates in GPCR Research GPCRLigNet: rapid screening for GPCR active ligands using machine learning. A robust approach for MicroED sample preparation of lipidic cubic phase embedded membrane protein crystals. Reviews, GPCRs, and more Cell-trafficking impairment in disease-associated LPA6 missense mutants and a potential pharmacoperone therapy for autosomal recessive woolly hair/hypotrichosis. Structural and Molecular Insights into GPCR Function Structure-based design of novel melanin-concentrating hormone receptor-1 ligands based on saturated nitrogen-containing heterocycles. Industry News Josephine (Pina) Cardarelli from GPCR Therapeutics on Beyond Biotech Podcast Crinetics Pharmaceuticals Reports Fourth Quarter And Full Year 2022 Financial Results And Provides Corporate Update Alastair Brown from Sosei Heptares, will present at the 18th Annual Biomarkers Congress. A growing understanding of the role of muscarinic receptors in the molecular pathology and treatment of schizophrenia Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023. GPCR Events, Meetings, and Webinars 2nd ERNEST Training School. (February 20 - March 3, 2023). WEBINAR Using integrative biophysical approaches to understand GPCR regulation by β-arrestins. (March 2, 2023) GEM2023. (March 14-17, 2023). SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). GPCR Jobs PhD Studentship - Integrated Approaches In Pharmacology, Computer Simulations And Machine Learning To Predict Ligand Signaling Pathways Via The Anti-Inflammatory Receptor Gpr84 PhD Studentship - Insight Into The Neuroprotective Function Of The Prostaglandin Receptor Ep2 Computational drug design PhD student at the Department of Drug Design and Pharmacology Computational drug design postdoc at the Department of Drug Design and Pharmacology Software / database development PhD student at Department of Drug Design and Pharmacology Software / database development postdoc at the Department of Drug Design and Pharmacology Postdoc in Bioinformatics/Data Science at Department of Drug Design and Pharmacology Scientist/Senior Scientist (Research Software Engineer) Senior Scientist – Native Mass Spectrometry Speculative Applications (Protein Biochemistry) Speculative Applications (Mass Spectrometry) Speculative Applications (Chemistry) Explore Dr. GPCR Ecosystem

Regulator G protein Signaling (RGS) proteins are critical components of the intracellular signaling pathways that mediate the effects of G protein-coupled receptors (GPCRs). Upon activation, GPCRs have conformational changes that allow the coupling and subsequent activation of the G-protein heterotrimeric complex (α, β, and γ); it is at this point when the RGS proteins play a key role in the deactivation of the alpha subunit contributing to the termination of the G protein-mediated signaling cascades[1, 2]. RGS proteins are a family with around 20 members characterized by the presence of a conserved RGS-homology (RH) domain. This domain contains the catalytic core that catalyzes the hydrolysis of guanosine triphosphate (GTP) to guanosine diphosphate of the G protein α subunit promoting the switch from activated to an inactivated state[3]. In addition to the RGS domain, RGS proteins also contain a range of other structural motifs that are critical for their function, including the G protein-binding domain, the DEP (Dishevelled, Egl-10 and Pleckstrin domain) domain, and the GoLoco motif[2, 3]. Role of RGS proteins in regulating GPCR signaling: Recent studies have revealed that the interaction between RGS proteins and GPCRs is mediated by a range of structural motifs, including the G protein-binding (GB) and the RGS domains. The interaction between RGS proteins and GPCRs is highly specific and tightly regulated; mutations in the RGS domain and other structural motifs have been shown to alter the specificity and potency of the RGS-GPCR interaction[2]. As negative regulators of GPCR signaling, RGS proteins play a critical role in regulating the duration and amplitude of GPCR signaling. For example, μ opioid receptor (MOR) interacts with Gαi/o and Gαz subunits, which have a slow enzymatic GTPase activity requiring the action of RGSs proteins. RGSs bind to GTP-bound Gα to accelerate GTP hydrolysis reducing the activity of the Gα subunit and resulting in negative regulation of MOR downstream signaling[3, 4]. Besides the differences in their structural complexity, some members of the RGS family are selective for certain GPCRs, as a proof RGS4 which is expressed in the brain, has been shown to modulate dopamine signaling by specifically regulating the activity of the dopamine D2 receptor; enhancing the activity of the G protein that is coupled to the receptor and leading to a decrease in dopamine signaling[4, 5]. Another signaling pathway related to RGS4 involves the regulation of the immune response. RGS4 is expressed in various immune cells, including T cells and B cells, and has been shown to modulate immune cell activation and cytokine production. RGS4 acts as a negative regulator of T cell activation, and its expression is upregulated in response to T cell activation[6]. Implications of RGS protein dysregulation in disease: The Dysregulation of RGS proteins has been implicated in a range of diseases, including cardiovascular disease, pain, hypertension, and cancer. In cardiovascular disease, RGS proteins play a critical role in regulating blood pressure and vascular function. Relating to pain, RGS4 in pain regulation is a topic of increasing interest because it has been identified as a key player in the modulation of nociception[7]. In hypertension, dysregulation of RGS proteins has been shown to contribute to the pathogenesis of the disease. While in cancer, RGS proteins are involved in regulating cell proliferation and survival[8]. In conclusion, RGS proteins are essential modulators for the GPCR signaling mediated by G proteins, which play a crucial role in regulating a range of physiological processes. The dysregulation of these proteins has been implicated in a range of diseases, and understanding the mechanisms of these complex molecules is crucial for developing effective therapies. 1. Tesmer, J.J., et al., Structure of RGS4 bound to AlF4--activated G(i alpha1): stabilization of the transition state for GTP hydrolysis. Cell, 1997. 89(2): p. 251-61. https://pubmed.ncbi.nlm.nih.gov/9108480/ 2. Senese, N.B., et al., Regulator of G-Protein Signaling (RGS) Protein Modulation of Opioid Receptor Signaling as a Potential Target for Pain Management. Front Mol Neurosci, 2020. 13: p. 5. https://pubmed.ncbi.nlm.nih.gov/32038168/ 3. Hollinger, S. and J.R. Hepler, Cellular regulation of RGS proteins: modulators and integrators of G protein signaling. Pharmacol Rev, 2002. 54(3): p. 527-59. https://pubmed.ncbi.nlm.nih.gov/12223533/ 4. Wang, Q., L.Y. Liu-Chen, and J.R. Traynor, Differential modulation of mu- and delta-opioid receptor agonists by endogenous RGS4 protein in SH-SY5Y cells. J Biol Chem, 2009. 284(27): p. 18357-67. https://pubmed.ncbi.nlm.nih.gov/19416973/ 5. Zhuang, Y., et al., Structural insights into the human D1 and D2 dopamine receptor signaling complexes. Cell, 2021. 184(4): p. 931-942.e18. https://pubmed.ncbi.nlm.nih.gov/33571431/ 6. Wang, D., The essential role of G protein-coupled receptor (GPCR) signaling in regulating T cell immunity. Immunopharmacol Immunotoxicol, 2018. 40(3): p. 187-192. https://pubmed.ncbi.nlm.nih.gov/29433403/ 7. Avrampou, K., et al., RGS4 Maintains Chronic Pain Symptoms in Rodent Models. J Neurosci, 2019. 39(42): p. 8291-8304. https://pubmed.ncbi.nlm.nih.gov/31308097/ 8. Hu, Y., et al., Identification of a five-gene signature of the RGS gene family with prognostic value in ovarian cancer. Genomics, 2021. 113(4): p. 2134-2144. https://pubmed.ncbi.nlm.nih.gov/33845140/

Hello Readers👋, Welcome to our GPCR weekly newsletter! Get ready to stay up-to-date with the latest research, advancements, and news in the field. Remember, you can always adjust your email preferences in your account settings. If you'd like to suggest papers to include in our news, please email them to us at Hello@DrGPCR.com. For Dr. GPCR News, please subscribe to our monthly newsletter. Below is your Classified GPCR News at a glance for February 13th to 19th, 2023. GPCR Activation and Signaling Statins inhibit protein kinase D (PKD) activation in intestinal cells and prevent PKD1-induced growth of murine enteroids. Anionic phospholipids control mechanisms of GPCR-G protein recognition. Mandibulofacial dysostosis with alopecia results from ETAR gain-of-function mutations via allosteric effects on ligand binding. From outside to inside and back again: the lysophosphatidic acid-CCN axis in signal transduction. Octopamine and tyramine signalling in Aedes aegypti: Molecular characterization and insight into potential physiological roles. The spatial distribution of GPCR and Gβγ activity across a cell dictates PIP3 dynamics. GPCR Binders, Drugs, and more Novel targets for potential therapeutic use in Diabetes mellitus. Reviews, GPCRs, and more Single nucleotide variations encoding missense mutations in G protein-coupled receptors may contribute to autism. Understanding Neuropeptide Transmission in the Brain by Optical Uncaging and Release. Structural and Molecular Insights into GPCR Function Quantitative analysis of sterol-modulated monomer-dimer equilibrium of the β1-adrenergic receptor by DEER spectroscopy. Industry News Trevena Announces Publication of OLINVYK Respiratory Physiology Study In Anesthesiology Neurocrine Biosciences discovers new muscarinic M4 receptor antagonists Alexander S. Hauser on receiving 4.8 mill DKK from Carlsberg Foundation’s Semper Ardens: Accelerate programme. From the scientist’s view: a conversation with … Chris Tate AI is dreaming up drugs that no one has ever seen. Now we've got to see if they work. Artificial Intelligence for Drug Discovery Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023. GPCR Events, Meetings, and Webinars SLAS2023 International Conference and Exhibition (February 25 - March 1, 2023). 2nd ERNEST Training School. (February 20 - March 3, 2023). WEBINAR Using integrative biophysical approaches to understand GPCR regulation by β-arrestins. (March 2, 2023) GEM2023. (March 14-17, 2023). SLAS 2023 B Building Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). GPCR Jobs Computational drug design PhD student at the Department of Drug Design and Pharmacology Computational drug design postdoc at the Department of Drug Design and Pharmacology Software / database development PhD student at Department of Drug Design and Pharmacology Software / database development postdoc at the Department of Drug Design and Pharmacology Postdoc in Bioinformatics/Data Science at Department of Drug Design and Pharmacology Scientist/Senior Scientist (Research Software Engineer) Senior Scientist – Native Mass Spectrometry Speculative Applications (Protein Biochemistry) Speculative Applications (Mass Spectrometry) Speculative Applications (Chemistry) Post-Doctoral Research Assistant Research Fellow - School of Physics and Astronomy Postdoctoral Research Associate (Fixed Term) Explore Dr. GPCR Ecosystem

Glycosylation and sulfation – N-terminal PTMs on chemokine receptors The interaction of chemokine receptors with their cognate chemokine ligands is generally described by the two-step/two-site model - the first step characterized by the interaction between the extracellular domains of the receptor and the structural core domain of the chemokine (CRS1); and the second step featured by the interaction between the N-terminus of the chemokine and the extracellular loops and transmembrane domains of the receptor (CRS2), which will trigger conformational changes that ultimately lead to receptor activation (Scholten D et al. 2012, Kufavera I et al. 2017). Within the CRS1 ineraction mode, the N-terminal region of chemokine receptors is indispensable for chemokine binding, where the negatively charged residues, as a consequence of the negatively charged aminoacids and post-translational modifications (PTMs), contribute to the high affinity binding to the positively charged groove on the chemokine. N-terminal PTMs include sulfation and glycosylation which contribute to the overall negative charge of the N-terminus fine-tuning chemokine binding. Chemokine receptors have either reported or predicted sites of N-acetyl galactosamine (GalNAc)-type O-glycosylation in their N-termini as well as sulfation, both PTMs which co-localize in the Trans-Golgi network (Mehta AY et al. 2020). In silico analyses done in this study with the NetOGlyc 4.0 prediction algorithm (Steentoft C et al. 2013) for O-glycosylation and the sulfinator tool for tyrosine sulfation sites (Monigatti F et al. 2002), suggest that CC chemokine receptors have general patterns of O-glycosylation and tyrosine sulfation in their N-terminal region which seem to be widely conserved between human and murine sequences. GalNAc-type or mucin-type O-glycosylation is initiated by the transfer of Gal-NAc to a serine or threonine residue by polypeptide GalNAc-transferases (GalNAc-Ts) which is further elongated with the linkage of other monosaccharides (Schjoldager KT et al. 2020), and usually capped by a single sialic acid, although, in rare cases glycans can carry polysialylation (PolySia) (Mindler K et al. 2021). PolySia has been described for CCR7 where it specifically affects the recognition of CCL21 but not CCL19, interfering with dendritic cell trafficking (Kiermaier E et al. 2016). Other examples of O-glycosylation impact on chemokine receptors include the viral receptor US28 (Bagdonaite I et al. 2016) where O-glycosylation contributes to the differential binding to CC or CX3C chemokine (Casarosa P et al. 2005). CCR5 O-glycosylation, which is under investigation in this study, also plays a major role in promoting the interaction with CCL3 and CCL5, being also important for HIV infection (Bannert N et al. 2001). Tyrosine sulfation consists in the transfer of a sulfate group from the adenosine 3’-phosphate 5’-phosphosulfate (PAPS) donor to the hydroxyl group of a tyrosine residue of the protein chain (Seibert C and Sakmar TP. 2008) by tyrosine sulfotransferase 1 or 2 (TPST1/2). This PTM has been shown to be heterogeneous [Li X et al. 2018; Scurci I et al. 2021) and to improve the affinity of chemokines through the charge interactions between the negative sulfate groups in the N-terminus and the positively charged chemokines (Ludeman JP and Stone MJ. 2014). CCR5 sulfation has been also reported to play a key role in CCL3 and CCL5 binding (Bannert N et al. 2001) as well as to exist in heterogeneous forms inside the cell (Scurci I et al. 2021). The atypical chemokine receptor 2 (ACKR2), US28 and sphingosine-1-phosphate receptor 1 (S1PR1) also carry tyrosine sulfation sites in their N-termini that mediate ligand binding and signaling (Bannert N et al. 2001; Casarosa P et al. 2005; Gao J et al. 2003; Fieger CB et al. 2005). Probing the modulation of O-glycosylation and tyrosine sulfation on CCR5 and CCR1 function In this study engineered cell lines, inhibitors and mutagenesis approaches were performed to evaluate the effects of O-glycosylation and tyrosine sulfation modulation on the chemokine receptors CCR1 and CCR5 pharmacology. Both PTMs were shown to contribute to the binding of CCL5 and CCL8 and to a minor extend CCL3. The interplay between these PTMs was also revealed where mutagenesis of tyrosine sulfation sites considerably impacted O-glycosylation. Removal of the terminal sialic acid was also shown to negatively impact signaling as previously appreciated (Bannert N et al. 2001). Interestingly, expression of PolySia was able to partially rescue the signaling upon reduction of sulfation, suggesting the fine-tuning role of O-glycosylation although the mechanism through which PolySia exerts its compensatory effect remains unclear. This effect could be driven by a specific carrier or it can potentially be related with the increase in the general negative charge of the cell surface glycosaminoglycans which have an established role in chemokine gradients and oligomerization (Deshauer C et al. 2015; Dyer DP et al. 2016). In this work authors set out to investigate GalNAc-Ts candidates involved in CCR5 O-glycosylation with CHO GalNAc-T knock-outs transfected with CCR5. From the five GalNAc-Ts, GalNAc-T1 was shown to be the most likely candidate for directly glycosylating CCR5 although T11 may also be involved. Sulfo-glyco barcode - perspectives and added value in drug discovery A growing body of evidence proposes that O-glycosylation and sulfation are important PTMs in chemokine receptor biology and pharmacology however the reported effects can vary depending on the receptor-ligand pairs and potentially cell line and tissue tested. The combined effects of both PTMs as well as the relevance of specific acceptor sites and glycan composition remains to be investigated in more depth. Dissecting the biological relevance of these PTMs requires relevant cells expressing the chemokine receptor endogenously together with relevant enzymes and co-receptor systems. In this study, although direct effects of O-glycosylation removal are ruled out it is possible that indirect effects may also contribute to the observed phenotype since many glycosyltransferases and the two TPSTs also carry O-glycosylation (King SL et al.2017). In addition, tyrosine sulfation is heterogenous between cell lines or even on the same cell (Scurci I et al. 2021). It remains to be discovered how much the modulation of these PTMs is relevant for future drug design. There are few examples of the applicability of the sulfo-glyco barcode in drug discovery. For CCR5, it has been shown that N-terminal antibodies show different sulfo-sensitivities (Scurci I et al. 2021). It has been also reported that PSGL-1 glycosulfo peptide analogue GSnP-6 displays nanomolar affinity and promising potential for blocking PSGL-1/P-selectin interaction (Wong DJ et al. 2021). Moreover, sulfated mCCR2 peptides can outcompete available chemokines, decreasing retinal degradation in mice (Jung SA et al. 2021). Further research will be needed to boost our understanding on the dynamics and biological relevance of these PTMs in the chemokine receptor system which may ultimately allow for new precise targeting of the immune system. Check the original article at https://pubmed.ncbi.nlm.nih.gov/36729338/ #GPCR #DrGPCR#Ecosystem

Hello Readers👋, We're excited to share our weekly newsletter with you. Keep up-to-date with the latest research and advancements in the field with our convenient, weekly news delivered to your inbox. You can always adjust your email preferences in your account settings. For Dr. GPCR News, please subscribe to our monthly newsletter. Below is your Classified GPCR News at a glance for February 6th to 12th, 2023. GPCR Activation and Signaling Ubiquitylation of BBSome is required for ciliary assembly and signaling. Isoform- and ligand-specific modulation of the adhesion GPCR ADGRL3/Latrophilin3 by a synthetic binder. Unravelling GPCR signalling networks using global phosphoproteomics. GPCR Binders, Drugs, and more An antibody-drug conjugate targeting GPR56 demonstrates efficacy in preclinical models of colorectal cancer. GPCRs in Neuroscience Heterotrimeric G proteins regulate planarian regeneration and behavior. Methods & Updates in GPCR Research Engineered Human Antibody with Improved Endothelin Receptor Type A Binding Affinity, Developability, and Serum Persistence Exhibits Excellent Antitumor Potency. Development and Characterization of a Highly Selective Turn-On Fluorescent Ligand for β3-Adrenergic Receptor. Quantitative analysis of sterol-modulated monomer-dimer equilibrium of the β1-adrenergic receptor by DEER spectroscopy. Industry News Biotech startup Structure pulls off rare IPO, raising $161M 3-drug combo leads to 'unprecedented' response in pancreatic cancer models Addex Therapeutics Ltd Strategic Partner Completes Enrollment in ADX71149 Epilepsy Phase 2 Study Part 1 IPO Raises $161M for GPCR-Based Drug Developer Structure Therapeutics Crinetics Pharmaceuticals Announces Inducement Grants Under Nasdaq Listing Rule 5635(c)(4) arcoscreen is one of the 13 biotech companies to discover in western in 2023 Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023. GPCR Events, Meetings, and Webinars 2nd GPCR-Targeted Drug Discovery Summit, (February 21 - 23, 2023), Boston. SLAS2023 International Conference and Exhibition (February 25 - March 1, 2023). 2nd ERNEST Training School. (February 20 - March 3, 2023). WEBINAR Using integrative biophysical approaches to understand GPCR regulation by β-arrestins. (March 2, 2023) GEM2023. (March 14-17, 2023). SLAS 2023 B Building Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). GPCR Jobs Director, Business Development & Strategy CADD and Informatics Head or Team Lead (depending on experience) with Structural Bioinformatics Expertise Fully Funded Doctoral Training Programme Studentships 2023 Computational drug design PhD student at the Department of Drug Design and Pharmacology Computational drug design postdoc at the Department of Drug Design and Pharmacology Software / database development PhD student at Department of Drug Design and Pharmacology Software / database development postdoc at the Department of Drug Design and Pharmacology Postdoc in Bioinformatics/Data Science at Department of Drug Design and Pharmacology Scientist/Senior Scientist (Research Software Engineer) Senior Scientist – Native Mass Spectrometry Speculative Applications (Protein Biochemistry) Speculative Applications (Mass Spectrometry) Speculative Applications (Chemistry) Post-Doctoral Research Assistant Research Fellow - School of Physics and Astronomy Postdoctoral Research Associate (Fixed Term) Explore Dr. GPCR Ecosystem

With around 187 people dying every day from an opioid overdose in the U.S., combatting the opioid overdose epidemic has become a mean challenge for the scientific community. According with the National Institute on Drug Abuse (NIDA) nearly 92, 000 Americans died from drug-involved overdose in 2020, of which approximately 75% involved opioids. Some examples of opioids include heroin, morphine, codeine, fentanyl, methadone, tramadol and other opioid analogs. Opioids are analgesic drugs consumed non-medically for euphoric feelings and medically for pain relief, although the last one comes with pharmacological side effects such as breathing difficulties and addiction which contribute to the current opioid crisis problem3. The opioid system is one of the most important in regulating the response to nociception, i.e. the response of the nervous system to painful stimuli; the 1970s marked the beginning of its study with the first discoveries which suggested that there were "binding sites" in the central nervous system that were recognized by exogenous opioids such as morphine, leading later to the discovery of opioid receptors1. The opioid system is composed of a set of major endogenous opioid peptides (EOPs): β-endorphin, enkephalins and dynorphins, and four opioid receptors (ORs): μ-opioid receptor (MOR), κ-opioid receptor (KOR), δ-opioid receptor (DOR) which are expressed throughout the central and peripheral nervous system and regulate important physiological functions such as analgesia, stress response, mood, reward, etc3. Opioid receptors belong to class A of G protein-coupled receptors or GPCRs and signaled mainly through Gai/o, beta/gamma subunits and arrestins. At the synapse, they are localized in both pre- and postsynaptic compartments and their activation is generally related to inhibiting neurotransmission by hyperpolarizing the cell or reducing or potentiating neuronal activity4. Given their pathophysiological significance in pain, addiction and depression opioid receptors represent important pharmacological targets. Morphine is one of the most widely used and proven analgesic for the treatment of severe acute or chronic pain conditions, but their use is overshadowed by their side effects and by the development of dependence and addiction. Morphine is an agonist of Mu opioid receptor (MOPR) and one of the objectives in the development of new opioids is to synthesize compounds with high analgesic power but without side effects like morphine does3. Therefore, the study of the different signal transductions triggered by the opioid-receptor interaction is of great importance. With this in mind one of the proposals is to take advantage of biased agonism, i.e. when the same receptor signals downstream through different signaling pathways triggered by different molecules2. There are several reports that have addressed the differences in the mechanisms of transduction triggered by MOR, reporting that G-protein signaling is more associated with the analgesic effect, while the side effects are orchestrated via β-arrestin 2. However, opioids that prevent recruiting β-arrestin 2 do not address the problem since ligands that only minimally recruit β-arrestin 2 to MORs may also cause opioid side effects2. Therefore to understand better the functions of arrestins in MOR signaling, Shiraki et al., explored the function of β-arrestin 2 in MOR signaling using the SH-SY5Y cell line that endogenously expresses MOR and was modified through CRISPR/Cas9 to knock out β-arrestin 1 and 2 gene expression. This report highlights a mechanism of β-arrestin pathway activation dependent on G protein activation, which contrasts with the idea that these signaling pathways are independent and compete with each other. The authors found that both β-arrestin 1 and 2 are involved in MOR internalization and downstream signaling activation of the β-arrestin pathway under Gi/o activation-MOR, being crucial the formation of β-arrestin/ β2-adaptin and clathrin heavy chain complex to mediates MAPK signaling5. These findings highlight how G proteins and β-arrestins are involved in driving intracellular signaling and reinforce the role of β-arrestins in the physiological opioid system. If you are interested in learning more about the molecular details of this study, you can consult the article at the following link https://pubmed.ncbi.nlm.nih.gov/36502633/ References 1. Brownstein MJ. A brief history of opiates, opioid peptides, and opioid receptors. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5391-3. doi: 10.1073/pnas.90.12.5391. PMID: 8390660; PMCID: PMC46725. https://pubmed.ncbi.nlm.nih.gov/35435616/ 2. Faouzi A, Varga BR, Majumdar S. Biased Opioid Ligands. Molecules. 2020 Sep 16;25(18):4257. doi: 10.3390/molecules25184257. PMID: 32948048; PMCID: PMC7570672. https://pubmed.ncbi.nlm.nih.gov/8390660/ 3. Manhapra A. Complex Persistent Opioid Dependence-an Opioid-induced Chronic Pain Syndrome. Curr Treat Options Oncol. 2022 Jul;23(7):921-935. doi: 10.1007/s11864-022-00985-x. Epub 2022 Apr 18. PMID: 35435616. https://pubmed.ncbi.nlm.nih.gov/35782382/ 4. Reeves KC, Shah N, Muñoz B, Atwood BK. Opioid Receptor-Mediated Regulation of Neurotransmission in the Brain. Front Mol Neurosci. 2022 Jun 15;15:919773. doi: 10.3389/fnmol.2022.919773. PMID: 35782382; PMCID: PMC9242007. https://pubmed.ncbi.nlm.nih.gov/32948048/ 5. Shiraki A, Shimizu S. The molecular associations in clathrin-coated pit regulate β-arrestin-mediated MAPK signaling downstream of μ-opioid receptor. Biochem Biophys Res Commun. 2023 Jan 15;640:64-72. doi: 10.1016/j.bbrc.2022.11.098. Epub 2022 Nov 30. PMID: 36502633. https://pubmed.ncbi.nlm.nih.gov/36502633/

Hello Readers👋, We're excited to share our weekly newsletter with you. Keep up-to-date with the latest research and advancements in the field with our convenient, weekly news delivered to your inbox. You can always adjust your email preferences in your account settings. For Dr. GPCR News, please subscribe to your monthly newsletter. Below is your Classified GPCR News at a glance for January 30th to February 5th, 2023. GPCR Activation and Signaling The dual-function chemokine receptor CCR2 drives migration and chemokine scavenging through distinct mechanisms. Cannabinoid 1 (CB1 ) receptor arrestin subtype-selectivity and phosphorylation dependence. Metabolic depletion of sphingolipids inhibits agonist-induced endocytosis of the serotonin1A receptor. Genetic code expansion to enable site-specific bioorthogonal labeling of functional G protein-coupled receptors in live cells. Platelet P2Y1 receptor exhibits constitutive G protein signaling and β-arrestin 2 recruitment. Multiple Potassium Channel Tetramerization Domain (KCTD) family members interact with Gβγ, with effects on cAMP signaling. GPCR Binders, Drugs, and more Ligands selectively tune the local and global motions of neurotensin receptor 1 (NTS1). Pharmacological characterization of novel small molecule agonists and antagonists for the orphan receptor GPR139. Valorization of leftover green tea residues through conversion to bioactive peptides using probiotics-aided anaerobic digestion. Comparative evaluation of biased agonists Sarcosine1 , d-Alanine8 -Angiotensin (Ang) II (SD Ang II) and Sarcosine1 , Isoleucine8 -Ang II (SI Ang II) and their radioiodinated congeners binding to rat liver membrane AT1 receptors. GPCRs in Cardiology, Endocrinology, and Taste Receptor autoantibodies: Associations with cardiac markers, histology, and function in human non-ischaemic heart failure. GPCRs in Oncology and Immunology LP2, a cyclic angiotensin-(1-7) analog extended with an N-terminal D-lysine, impairs growth of patient-derived xenografts of colorectal carcinoma in mice. Simultaneous activation of CXC chemokine receptor 4 and histamine receptor H1 enhances calcium signaling and cancer cell migration. Methods & Updates in GPCR Research Deep-Learning-Enhanced Diffusion Imaging Assay for Resolving Local-Density Effects on Membrane Receptors. Computational investigation of functional water molecules in GPCRs bound to G protein or arrestin. Functional expression of oxytocin receptors in pulp-dentin complex. Reviews, GPCRs, and more Understanding Neuropeptide Transmission in the Brain by Optical Uncaging and Release. Patching holes in the mechanism of opioid tolerance. Non-canonical Golgi-compartmentalized Gβγ signaling: mechanisms, functions, and therapeutic targets. β-arrestins and G protein-coupled receptor kinases in viral entry: A graphical review. Development and challenges in the discovery of 5-HT1A and 5-HT7 receptor ligands. Involvement of SAP97 anchored multiprotein complexes in regulating cardiorenal signaling and trafficking networks. The chemokines CXCL8 and CXCL12: molecular and functional properties, role in disease and efforts towards pharmacological intervention. The Memory Orchestra: Contribution of Astrocytes. Structural and Molecular Insights into GPCR Function Structural view of G protein-coupled receptor signaling in the retinal rod outer segment. Cryo-EM structure of orphan G protein-coupled receptor GPR21. Industry News Exscientia Announces First-in-Human Study for Bristol Myers Squibb In-Licensed PKC Theta Inhibitor, EXS4318 GPCR Therapeutics Announces Launch of US Phase 2 Trial of GPC-100 in Multiple Myeloma Behind Friday's $161M IPO: A star scientist, GPCR drug discovery and a plan to challenge pharma in diabetes TGA’s approval of MDMA and Psilocybin for medical use is a result of ‘renewed interest’ Addex Strategic Partner Completes Enrollment In Adx71149 Epilepsy Phase 2 Study Part 1 Domain Therapeutics to Participate in Upcoming Investor and Industry Conferences International Day of Women and Girls in Science Orion Biotechnology Publishes Whitepaper Highlighting the Importance and Challenges in Unlocking “Undruggable” GPCRs Endocrine Society Awards Baxter Prize to Innovator in Endocrine Drug Discovery Upstream Bio Announces the Appointment of Marcella Ruddy, M.D., to the Board of Directors Call for GPCR Papers GPCRs: Signal Transduction. Deadline extended to March 31st, 2023 GPCR Events, Meetings, and Webinars 2nd GPCR-Targeted Drug Discovery Summit, (February 21 - 23, 2023), Boston. SLAS2023 International Conference and Exhibition (February 25 - March 1, 2023). 2nd ERNEST Training School. (February 20 - March 3, 2023). WEBINAR Using integrative biophysical approaches to understand GPCR regulation by β-arrestins. (March 2, 2023) GEM2023. (March 14-17, 2023). SLAS 2023 B uilding Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). GPCR Jobs Computational drug design PhD student at the Department of Drug Design and Pharmacology Computational drug design postdoc at the Department of Drug Design and Pharmacology Software / database development PhD student at Department of Drug Design and Pharmacology Software / database development postdoc at the Department of Drug Design and Pharmacology Postdoc in Bioinformatics/Data Science at Department of Drug Design and Pharmacology Scientist/Senior Scientist (Research Software Engineer) Senior Scientist – Native Mass Spectrometry Speculative Applications (Protein Biochemistry) Speculative Applications (Mass Spectrometry) Speculative Applications (Chemistry) Post-Doctoral Research Assistant Research Fellow - School of Physics and Astronomy Postdoctoral Research Associate (Fixed Term) Explore Dr. GPCR Ecosystem

Hello Readers👋, We're excited to share our weekly newsletter with you. Keep up-to-date with the latest research and advancements in the field with our convenient, weekly news delivered to your inbox. You can always adjust your email preferences in your account settings Below is your Classified GPCR News at a glance for January 23rd to the 29th, 2023. GPCR Activation and Signaling A multi-dimensional view of context-dependent G protein-coupled receptor function. The role of intracellular calcium and Rho kinase pathways in G protein-coupled receptor-mediated contractions of urinary bladder urothelium and lamina propria. Soluble cyclase-mediated nuclear cAMP synthesis is sufficient for cell proliferation. Molecular Modeling Study of a Receptor-Orthosteric Ligand-Allosteric Modulator Signaling Complex. GPCR Binders, Drugs, and more MRAP2 regulates energy homeostasis by promoting primary cilia localization of MC4R. Design, synthesis, and evaluation of substituted alkylindoles that activate G protein-coupled receptors distinct from the cannabinoid CB1 and CB2 receptors. Methods & Updates in GPCR Research Development and Characterization of a Highly Selective Turn-On Fluorescent Ligand for β3-Adrenergic Receptor. GPCR Signaling Measurement and Drug Profiling with an Automated Live-Cell Microscopy System. Generation of Gαi knock-out HEK293 cells illuminates Gαi-coupling diversity of GPCRs. Reviews, GPCRs, and more Pharmacological Profiling of a Brugia malayi Muscarinic Acetylcholine Receptor as a Putative Antiparasitic Target. Structural and Molecular Insights into GPCR Function Structural and dynamic insights into supra-physiological activation and allosteric modulation of a muscarinic acetylcholine receptor. Structural Understanding of Peptide-Bound G Protein-Coupled Receptors: Peptide-Target Interactions. Structural and Molecular Determinants for Isoform Bias at Human Histamine H3 Receptor Isoforms. Industry News Inversago Pharma to Participate at the 2023 SVB Securities Global BioPharma Conference Orbit Discovery and Endevica Bio enter multi-target collaboration to advance development of cachexia therapeutics Sosei Heptares Webinar Presentation for FY2022 Financial Results Call for GPCR Papers GPCRs: Signal Transduction. Deadline February 12, 2023 GPCR Events, Meetings, and Webinars 2nd GPCR-Targeted Drug Discovery Summit, (February 21 - 23, 2023), Boston. SLAS2023 International Conference and Exhibition (February 25 - March 1, 2023). 2nd ERNEST Training School. (February 20 - March 3, 2023). GEM2023. (March 14-17, 2023). SLAS 2023 Building Biology in 3D Symposium. (April 20 - 21, 2023) SLAS Europe 2023 Conference and Exhibition. (May 22 - 26, 2023) 2nd LEAPS Meets Life Sciences Conference. (May 14 - 19, 2023) 8th and final ERNEST Meeting in Crete. (May 3 - 7, 2023). The Illuminating the Understudied Druggable Proteome Conference. (June 4 - 8, 2023). 2023 Molecular Pharmacology (GRS) Seminar GRC. (June 10 - 11, 2023). Progressive Technologies and Approaches Revealing Novel GPCR Biology and Drug Development Potential. (June 11 - 16, 2023). 19th World Congress of Basic & Clinical Pharmacology 2023. (July 2 - 7, 2023). GPCR Jobs Scientist/Senior Scientist (Research Software Engineer) Senior Scientist – Native Mass Spectrometry Speculative Applications (Protein Biochemistry) Speculative Applications (Mass Spectrometry) Speculative Applications (Chemistry) Post-Doctoral Research Assistant Research Fellow - School of Physics and Astronomy Postdoctoral Research Associate (Fixed Term) PhD Project - Investigating the lipid regulation of GPCR signalling Postdoctoral Fellow - Molecular Dynamics Postdoc in lipid regulation of GPCRs Postdoctoral positions, biophysics of transporters and GPCRs Chief of Staff Postdoctoral Fellow, Biochemical And Cellular Pharmacology Project Leader, Biology Explore Dr. GPCR Ecosystem

Executive Summary This whitepaper will provide an overview of G Protein-Coupled Receptors (GPCRs) and discuss current trends in GPCR drug discovery. GPCRs are an important class of drug targets that represent approximately 30% of global drug market sales. However, the receptors that these medicines target have been described as the ‘low-hanging’ fruit, and many of the remaining GPCRs have shown low tractability using established drug discovery approaches. This situation is now evolving, with recent scientific and technological breakthroughs encouraging a new wave of GPCR drug discovery. Among the challenging GPCRs are approximately 50 that have large natural ligands. While these small protein GPCRs are valuable drug targets linked to serious diseases, many remain undrugged because they are less tractable to standard drug discovery approaches. To solve this industry-wide problem, Orion Biotechnology (Orion) is introducing a novel solution driven by its PROcisionXᵀᴹ platform: precision-engineering analogs based on the natural small protein ligands of these receptors to unlock their therapeutic potential. The GPCR Opportunity GPCRs are the largest and most diverse group of membrane receptors. They are composed of seven membrane-spanning α-helices, comprising the transmembrane (TM) domain of the receptor, joined by alternating intracellular and extracellular loop regions that form the intracellular and extracellular faces of the receptor. In general, signaling occurs when agonists engage key microswitch structures located in the TM domain. These interactions induce local conformational changes which are amplified and transmitted to the intracellular face of the receptor, leading to the binding and activation of cytosolic effector proteins: the G proteins after which GPCRs are named, and arrestins which both shut off G protein activity and elicit G protein-independent signaling pathways. There are approximately 800 human GPCRs in the superfamily, controlling a broad range of physiological activities. Roughly half of the receptors in the superfamily are sensory GPCRs, involved in olfaction, taste, vision and pheromone signaling. The remaining non-sensory receptors, with functions more readily linked to pathology, are currently the main focus for GPCR drug discovery. A New Wave of GPCR Drug Discovery GPCRs are considered highly druggable, with GPCR-targeting therapeutics contributing ~30% of global drug market sales. However only ~15% of the GPCR superfamily has been successfully drugged, due in part to the intractability of certain groups within the GPCR superfamily to standard drug discovery approaches. Spurred on by new scientific and technological advances related to the understanding of structure and activation mechanisms of GPCRs, the biotech investment community has injected significant funding into small biotech companies developing novel technology platforms to target GPCRs in recent years. Within the last two years, three investments stand out: · Tectonic Therapeutics: received $80 Million USD in Series A financing to develop GPCR-targeting nanobodies. · Septerna: closed a $100 Million USD Series A round to develop small molecule drugs against difficult-to-drug GPCRs. · Domain Therapeutics: completed a $42 Million USD Series A round to develop novel immunotherapies, including an anti-CCR8 antibody. Small Protein GPCRs are Difficult to Drug Among the GPCRs that are challenging to drug using standard approaches is a group of approximately 50 receptors whose endogenous ligands are small proteins. These receptors have much larger binding pockets than the majority of GPCRs that have been successfully drugged so far. The significantly increased surface area of ligand-receptor interface for these GPCRs makes modulation with small molecule drugs, when they can be identified, much more challenging. Firstly, small molecule inhibitors only occupy a fraction of the available surface area in the large binding pocket, making them unsuitable for achieving full blockade of the natural ligand. Secondly, because the activation mechanism of small protein GPCRs involves making key contacts with the receptor at structurally distant sites in the binding pocket, it is difficult to achieve a high level of control over the quantity and quality of signaling activity using small molecule agonists. Significant resources have been invested into the search for small molecule modulators of small protein GPCRs, but discovery is challenging and the attrition rate during clinical development has been very high. Developing Small Protein Therapeutics with a Novel Discovery Platform One way to overcome the challenges presented by small protein GPCRs is to use small proteins as modalities to target them. These approaches include generating ligand analogs by precision engineering the natural small protein ligand of the receptor. The natural ligand of the target receptor provides an excellent scaffold, already with a size and shape that matches its binding pocket. The engineered analog approach involves optimizing this scaffold, adding and subtracting molecular contacts between the ligand and the receptor to obtain the required potency and signaling output. Small protein GPCRs feature a two-component binding mechanism in which one part of the ligand engages the extracellular face of the receptor, providing an ‘address’ function: binding affinity and specificity. The other part of the ligand reaches in to contact the TM domain of the receptor, providing a ‘message’ function: engaging the microswitches that control signaling activity. As seen below in Figure 1, natural ligands do not make use of all the potential binding contacts with the TM domain of the receptor, and by modifying these key points of contact it is possible not only to strongly increase binding affinity but also to fine tune both the quantity and quality of receptor signaling. Figure 1. Left: Natural ligands of small protein GPCRs use a two-component binding mechanism, with extracellular domain binding providing a highly specific ‘address’ function, and the transmembrane domain binding making ‘message’ contacts that drive signaling activity. Right: Orion ligand analogs are engineered to optimize transmembrane domain providing significantly increased binding affinity and providing a means to control both the quantity and quality of signaling activity. Orion’s approach to engineering ligand analogs involves leaving the ‘address’ binding interface on the extracellular face of the receptor unchanged and searching for optimized interaction with the TM domain of the receptor. This characteristic provides a major advantage in drug discovery: massive parallel library-based screening can be carried out directly on living cells expressing the target receptor. The unmodified ‘address’ interaction ensures that all molecules in the library are specific for the target receptor rather than irrelevant cell surface targets, meaning that in contrast to all of the other current GPCR discovery procedures, Orion’s approach does not require the onerous and time-consuming processes of target receptor expression, purification and stabilization at the beginning of a discovery campaign. Orion’s PROcisionXᵀᴹ Platform In Orion’s discovery process, shape space at the TM domain ‘message’ site is extensively explored by using surface display technology, with molecular diversity focused into the part of the ligand that engages the TM domain. Libraries of billions of analogs are screened in parallel for functional interaction with receptors presented in their physiological environment. Selection of the libraries typically leads to the isolation of several hundred candidate hits, enriched because they either bind with increased affinity at the cell surface, or because they have an enhanced capacity to elicit receptor internalization, thereby gaining the capacity to shelter inside the cell. Both properties enable enhanced ligands to avoid being eliminated during the stringent washing process. All candidate hits are then rapidly produced in parallel by total chemical synthesis using Orion’s proprietary multiplex synthesis technology, and then screened for pharmacological activity (receptor binding affinity, G protein and arrestin signaling) in high throughput cell-based assays. Screening directly identifies leads, and the structure-activity data obtained for the ensemble of candidate hits provides a rich resource of information to inform lead optimization, either by human and in silico-aided rational design, or by the construction of next-generation surface display libraries. The PROcisionXᵀᴹ platform (summarized in Figure 2) can be used to complete a discovery campaign within only 6-12 months, yielding optimized leads ready for preclinical validation. Hence PROcisionXᵀᴹ is one of the fastest discovery platforms in industry, and one that is uniquely capable of generating drug candidates with optimal molecular pharmacological properties for targeting small protein GPCRs. Figure 2: Workflow in Orion’s PROcisionXᵀᴹ platform. A unique and important feature of Orion’s discovery process is that the initial search for potent target receptor modulators is agnostic towards signaling activity because it selects for any kind of enhanced functional interaction with the receptor in the environment of a living cell. Since shape space exploration is specifically focused on the ‘message’ site in the TM domain, this means that lead candidates across the whole spectrum of signaling activity are identified: full antagonists, partial agonists with different levels of signaling activity, biased agonists that preferentially activate either G protein signaling or arrestin signaling, and superagonists with signaling activity higher than that of the native ligand (Figure 3). Figure 3: In a single discovery campaign (CCR5 is shown as an example), Orion’s PROcisionXᵀᴹ platform generates a diverse set of potent small protein ligand analogs across the whole spectrum of signaling activity. Orion has demonstrated that optimized antagonist analogs generated using the PROcisionXᵀᴹ platform have best-in-class in vitro functional inhibitory potency and show remarkably persistent binding. This leads to strikingly long in vitro receptor occupancy durations (at least seven days), meaning that lengthy pharmacodynamic activity is attained in vivo, despite the short circulatory half-life that is expected with small protein drugs. Orion’s receptor antagonist analogs have demonstrated powerful efficacies across a range of animal models spanning different disease indications. Orion’s superagonist analogs are not only powerful candidate drugs for receptor modulation in indications where potent receptor activation is required, they are also highly effective vehicles for payload delivery. This is due to their enhanced capacity to drive receptor internalization in a process that pulls the ligand, together with its conjugated payload, inside the target cell. Orion has demonstrated that this target cell-specific delivery cannot be achieved using the native ligand of the receptor, despite its modest level of receptor internalization activity (Figure 4). Figure 4: Payload delivery using Orion’s superagonist analogs on target cells expressing CCR5. The cytotoxic drug Monomethyl auristatin E (MMAE) was used alone or conjugated to either the native CCL5 ligand or to an Orion CCL5 superagonist analog. While the native ligand conjugate exhibited poor selectivity for CCR5-expressing cells and was less effective at killing cells than unconjugated MMAE, the Orion CCL5 superagonist analog significantly increased both the potency and selectivity of the payload towards target cells. Case study: Discovering a CCR2 Antagonist The chemokine receptor CCR2 regulates the recruitment of monocytes to the sites of inflammation. Since many inflammatory diseases are driven by inappropriate recruitment of monocytes into tissues, CCR2 has been described as the master controller of inflammatory pathology and is considered a valuable drug target for a number of inflammatory diseases including atherosclerosis, scleroderma, multiple sclerosis and cancer. Orion’s CCR2 antagonist analog (OB-004), discovered in only 6 months, is considerably more potent than the leading small molecule competitors in in vitro functional inhibition assays (Figure 5). In an ex vivo human endothelial transmigration model, OB-004 demonstrated an unprecedented level of monocyte blockade, robustly outperforming the most potent of the competitor small molecules (Figure 6). Orion used its discovery platform technology to rapidly develop a murinized version of OB-004 that was evaluated in a thioglycolate-induced peritonitis model. In this model, OB-004 showed powerful efficacy, achieving full blockade of monocyte recruitment in response to the inflammatory challenge (Figure 7). Figure 5: In an in vitro functional signaling assay on a human monocytic cell line, OB-004 demonstrated best-in-class potency versus a group of small molecule CCR2 inhibitors in clinical development. Figure 6: In a flow-based human monocyte endothelial transmigration assay, OB-004 strongly outperformed BMS813160 at all three concentrations tested, achieving full blockade of transmigration at the highest concentration in this stringent model. Figure 7: In vivo efficacy of murinized OB-004 (mOB-004) in the thioglycolate (TG) induced peritonitis murine model. Twice-daily treatment with mOB-004 dose-dependently suppressed monocyte infiltration into the peritoneum cavity, achieving complete blockade at the highest dose level. The Future of GPCR Research GPCRs are a very important class of drug targets, but despite historical successes, the majority of the superfamily remains undrugged, in part due to limitations in current drug discovery technology. There is a great deal of scientific and business interest in solving this problem, and the last two years have seen the emergence of a new wave of technology driven GPCR drug discovery companies. Available structural data across the GPCR superfamily is increasing at an exponential rate, alongside understanding of the mechanisms underlying receptor activation. This, together with the rapid development of artificial intelligence (AI)-driven in silico-based molecular interaction prediction, will inevitably lead to a more and more central role for in silico methodology and AI in GPCR drug discovery. Orion has entered a strategic research collaboration with Peptilogics, a US-based AI drug discovery company to exploit the rewards that can be reaped at this exciting new frontier. GPCRs with small protein ligands are among the receptor groups that have proven to be particularly challenging, and Orion has developed a unique and powerful solution to the problem, based on using the natural ligands of target receptors to generate precision-engineering analogs. Orion’s PROcisionXᵀᴹ platform has been validated by rapid success, obtaining not only best-in-class antagonists with strong in vivo efficacy, but also potent signaling molecules for indications where subtle tuning of receptor signaling is required, or as vehicles for payload delivery. In this way Orion has established itself as one of the leaders in the exciting new wave of GPCR drug discovery. About Orion Biotechnology Orion’s mission is to unlock the therapeutic potential of previously undruggable GPCRs. Orion has world-renowned expertise in GPCR pharmacology and protein engineering, and its proprietary drug discovery platform (PROcisionXᵀᴹ) has been used to rapidly and efficiently advance a diversified portfolio of GPCR-targeted drug candidates for the treatment of cancer and other serious diseases. Orion’s objectives include expanding internal and external pipelines and continuing to innovate technologies to unlock GPCRs. For more information, follow Orion Biotechnology on LinkedIn or visit www.orionbiotechnology.com. Relevant Publications Development of Orion’s platform technology Hartley, O. et al. (2004) Proceedings of the National Academy of Sciences of the United States of America, 101, 16460-16465. https://doi.org/10.1073/pnas.0404802101 · Initial work on precision engineering of a small protein GPCR ligand using new peptide chemistry technology and rational design Gaertner, H. et al. (2008). Proceedings of the National Academy of Sciences, 105(46), 17706–17711. https://doi.org/10.1073/pnas.0805098105 · Development and use of new technology to gain full control of signaling activity through a small protein GPCR via library-based screening Dorgham, K. et al. (2016) Methods in Enzymology, 570, 47-72. https://doi.org/10.1016/bs.mie.2015.09.014 · Library-based screening methodology for engineering small protein GPCR ligands Paolini-Bertrand, M. et al. (2018) The Journal of Biological Chemistry, 293(49), 19092–19100. https://doi.org/10.1074/jbc.RA118.004370 · New technology to rapidly synthesize candidate hits from library-based screening so that they can be tested in cell-based assays Akondi, K. B. et al. (2021) Chimia, 75(6), 489–494. https://doi.org/10.2533/chimia.2021.489 · Description of the fully integrated PROcisionXᵀᴹ discovery platform Structure-based validation of Orion’s approach Zheng, Y. et al. (2017) Immunity, 46, 1005-1017.e1005. https://doi.org/10.1016/j.immuni.2017.05.002 · Structural explanation of the receptor binding mechanism of a highly potent antagonist analog Isaikina, P. et al. (2021). Sci Adv, 7. https://doi.org/10.1126/sciadv.abg8685 · Structural explanation of the binding and activation mechanism of a highly potent superagonist analog In vivo efficacy validation of Orion’s optimized leads Lederman, M.M. et al. (2004) Science, 306, 485–487. https://doi.org/10.1126/science.1099288 · Use of a first precision-engineered analog to validate topical inhibition of CCR5 as a strategy for HIV prevention Veazey, R.S. et al. Journal of Infectious Diseases, 199, 1525-1527. https://doi.org/10.1086/598685 · Demonstration that topically administered precision engineered small proteins show full in vivo efficacy in a highly stringent model of HIV transmission Steinbach, K. et al. (2019) Science Translational Medicine, 11. https://doi.org/10.1126/scitranslmed.aav5519 · Demonstration of the in vivo efficacy of a systemically administered antagonist analog in a model of neuroinflammation Feasibility validation of Orion analogs in clinical development Cerini, F. et al. (2016) Protein Expression and Purification, 119, 1-10. https://doi.org/10.1016/j.pep.2015.10.011 · Demonstration of the feasibility of manufacturing clinical grade small protein analogs McGowan, I.M. et al. (2021). AIDS Res Hum Retroviruses, 37, 453-460. https://doi.org/10.1089/aid.2021.0010 · Successful first-in-human clinical study of a topically administered small protein analog The whitepaper can also be accessed at: https://fb-resources.fiercebiotech.com/free/w_defa3729/prgm.cgich=WP-Orion-02012023-WP