Search Results

October 2022 High hedgehog signaling is transduced by a multikinase-dependent switch controlling the apico-basal distribution of the GPCR smoothened "The oncogenic G-protein-coupled receptor (GPCR) Smoothened (SMO) is a key transducer of the hedgehog (HH) morphogen, which plays an essential role in the patterning of epithelial structures. Here, we examine how HH controls SMO subcellular localization and activity in a polarized epithelium using the Drosophila wing imaginal disc as a model. We provide evidence that HH promotes the stabilization of SMO by switching its fate after endocytosis toward recycling. This effect involves the sequential and additive action of protein kinase A, casein kinase I, and the Fused (FU) kinase. Moreover, in the presence of very high levels of HH, the second effect of FU leads to the local enrichment of SMO in the most basal domain of the cell membrane. Together, these results link the morphogenetic effects of HH to the apico-basal distribution of SMO and provide a novel mechanism for the regulation of a GPCR." Read more at the source #DrGPCR #GPCR #IndustryNews

October 2022 "We describe production of the human A2A adenosine receptor (A2AAR), a class A G protein-coupled receptor (GPCR) for 19F-NMR and single-molecule fluorescence (SMF) spectroscopy. We explain in detail steps shared between the two sample preparation strategies, including expression and isolation of A2AAR and assembly of A2AAR in lipid nanodiscs and procedures for incorporation of either 19F-NMR or fluorescence probes. Protocols for SMF experiments include sample setup, data acquisition, data processing, and error analysis. For complete details on the use and execution of this protocol, please refer to Wei et al. (2022) and Sušac et al. (2018)." Read more at the source #DrGPCR #GPCR #IndustryNews

October 2022 Regulation of pulmonary surfactant by the adhesion GPCR GPR116/ADGRF5 requires a tethered agonist-mediated activation mechanism "The mechanistic details of the tethered agonist mode of activation for the adhesion GPCR ADGRF5/GPR116 have not been completely deciphered. We set out to investigate the physiological importance of autocatalytic cleavage upstream of the agonistic peptide sequence, an event necessary for NTF displacement and subsequent receptor activation. To examine this hypothesis, we characterized tethered agonist-mediated activation of GPR116 in vitro and in vivo. A knock-in mouse expressing a non-cleavable GPR116 mutant phenocopies the pulmonary phenotype of GPR116 knock-out mice, demonstrating that tethered agonist-mediated receptor activation is indispensable for function in vivo. Using site-directed mutagenesis and species-swapping approaches, we identified key conserved amino acids for GPR116 activation in the tethered agonist sequence and in extracellular loops 2/3 (ECL2/3). We further highlight residues in transmembrane 7 (TM7) that mediate stronger signaling in mouse versus human GPR116 and recapitulate these findings in a model supporting tethered agonist:ECL2 interactions for GPR116 activation." Read more at the source #DrGPCR #GPCR #IndustryNews

August 2022 A correlation study of adhesion G protein-coupled receptors as potential therapeutic targets in Uterine Corpus Endometrial cancer "Adhesion G protein-coupled receptors (adhesion GPCRs), as a member of the G protein-coupled receptors (GPCRs) superfamily, have gradually entered the field of vision of researchers. The structure, function, and involvement of adhesion GPCRs in cancer development have been discussed in a series of papers. Uterine Corpus Endometrial Carcinoma (UCEC) isa malignanttumorofendometrium epithelial, whichis alsooneofthemostcommonfemalereproductivesystemtumors, but there are few pieces of research related to adhesion GPCRs in UCEC." Read more at the source #DrGPCR #GPCR #IndustryNews

September 2022 Fly casting with ligand sliding and orientational selection supporting complex formation of a GPCR and a middle sized flexible molecule "A GA-guided multidimensional virtual-system coupled molecular dynamics (GA-mD-VcMD) simulation was conducted to elucidate binding mechanisms of a middle-sized flexible molecule, bosentan, to a GPCR protein, human endothelin receptor type B (hETB). GA-mD-VcMD is a generalized ensemble method that produces a free-energy landscape of the ligand-receptor binding by searching large-scale motions accompanied with stable maintenance of the fragile cell-membrane structure. All molecular components (bosentan, hETB, membrane, and solvent) were represented with an all-atom model. Then sampling was conducted from conformations where bosentan was distant from the binding site in the hETB binding pocket. The deepest basin in the resultant free-energy landscape was assigned to native-like complex conformation. The following binding mechanism was inferred. First, bosentan fluctuating randomly in solution is captured using a tip region of the flexible N-terminal tail of hETB via nonspecific attractive interactions (fly casting). Bosentan then slides occasionally from the tip to the root of the N-terminal tail (ligand–sliding). During this sliding, bosentan passes the gate of the binding pocket from outside to inside of the pocket with an accompanying rapid reduction of the molecular orientational variety of bosentan (orientational selection). Last, in the pocket, ligand–receptor attractive native contacts are formed. Eventually, the native-like complex is completed. The bosentan-captured conformations by the tip-region and root-region of the N-terminal tail correspond to two basins in the free-energy landscape. The ligand-sliding corresponds to overcoming of a free-energy barrier between the basins." Read more at the source #DrGPCR #GPCR #IndustryNews

September 2022 "Research on cancer therapies focuses on processes such as angiogenesis, cell signaling, stemness, metastasis, and drug resistance and inflammation, all of which are influenced by the cellular and molecular microenvironment of the tumor. Different strategies, such as antibodies, small chemicals, hormones, cytokines, and, recently, gene editing techniques, have been tested to reduce the malignancy and generate a harmful microenvironment for the tumor. Few therapeutic agents have shown benefits when administered alone, but a few more have demonstrated clear improvement when administered in combination with other therapeutic molecules. In 2008 (and for the first time in the clinic), the therapeutic benefits of the β-adrenergic receptor antagonist, propranolol, were described in benign tumors, such as infantile hemangioma. Propranolol, initially prescribed for high blood pressure, irregular heart rate, essential tremor, and anxiety, has shown, in the last decade, increasing evidence of its antitumoral properties in more than a dozen different types of cancer. Moreover, the use of propranolol in combination therapies with other drugs has shown synergistic antitumor effects. This review highlights the clinical trials in which propranolol is taking part as adjuvant therapy at single administration or in combinatorial human trials, arising as a good pick and roll partner in anticancer strategies." Read more at the source #DrGPCR #GPCR #IndustryNews

September 2022 "The follicle-stimulating hormone receptor (FSHR) belongs to the glycoprotein hormone receptors, a subfamily of G-protein-coupled receptors (GPCRs). FSHR is involved in reproductive processes such as gonadal development and maturation. Structurally, the extensive extracellular domain, which contains the hormone-binding site and is linked to the transmembrane domain by the hinge region (HR), is characteristic for these receptors. How this HR is involved in hormone binding and signal transduction is still an open question. We combined in vitro and in situ chemical crosslinking, disulfide pattern analysis, and mutation data with molecular modeling to generate experimentally driven full-length models. These models provide insights into the interface, important side-chain interactions, and activation mechanism. The interface indicates a strong involvement of the connecting loop. A major rearrangement of the HR seems implausible due to the tight arrangement and fixation by disulfide bonds. The models are expected to allow for testable hypotheses about signal transduction and drug development for GPHRs." Read more at the source #DrGPCR #GPCR #IndustryNews

September 2022 "Background Spinal cord injury (SCI) is a common trauma in clinical practices. Subacute SCI is mainly characterized by neuronal apoptosis, axonal demyelination, Wallerian degeneration, axonal remodeling, and glial scar formation. It has been discovered in recent years that inflammatory responses are particularly important in subacute SCI. However, the mechanisms mediating inflammation are not completely clear. Methods The gene expression profiles of GSE20907, GSE45006, and GSE45550 were downloaded from the GEO database. The models of the three gene expression profiles were all for SCI to the thoracic segment of the rat. The differentially expressed genes (DEGs) and weighted correlation network analysis (WGCNA) were performed using R software, and functional enrichment analysis and protein–protein interaction (PPI) network were performed using Metascape. Module analysis was performed using Cytoscape. Finally, the relative mRNA expression level of central genes was verified by RT-PCR." Read more at the source #DrGPCR #GPCR #IndustryNews

September 2022 "Metabotropic γ-aminobutyric acid receptor (GABABR), a class C G protein-coupled receptor (GPCR) heterodimer, plays a crucial role in the central nervous system. Cryo-electron microscopy studies revealed a drastic conformational change upon activation and a unique G protein (GP) binding mode. However, little is known about the mechanism for GP coupling and activation for class C GPCRs. Here, we use molecular metadynamics computations to predict the mechanism by which the inactive GP induces conformational changes in the GABABR transmembrane domain (TMD) to form an intermediate pre-activated state. We find that the inactive GP first interacts with TM3, which further leads to the TMD rearrangement and deeper insertion of the α5 helix that causes the Gα subunit to open, releasing GDP, and forming the experimentally observed activated structure. This mechanism provides fresh insights into the mechanistic details of class C GPCRs activation expected to be useful for designing selective agonists and antagonists." Read more at the source #DrGPCR #GPCR #IndustryNews

G protein-coupled receptors (GPCRs) transmit extracellular signals to the inside by activation of intracellular effector proteins. Different agonists can promote differential receptor-induced signaling responses - termed bias - potentially by eliciting different levels of recruitment of effector proteins. As activation and recruitment of effector proteins might influence each other, thorough analysis of bias is difficult. Here, we compared the efficacy of seven agonists to induce G protein, G protein-coupled receptor kinase 2 (GRK2), as well as arrestin3 binding to the muscarinic acetylcholine receptor M3 by utilizing FRET-based assays. In order to avoid interference between these interactions, we studied GRK2 binding in the presence of inhibitors of Gi and Gq proteins and analyzed arrestin3 binding to prestimulated M3 receptors to avoid differences in receptor phosphorylation influencing arrestin recruitment. We measured substantial differences in the agonist efficacies to induce M3R-arrestin3 versus M3R-GRK2 interaction. However, the rank order of the agonists for G protein- and GRK2-M3R interaction was the same, suggesting that G protein and GRK2 binding to M3R requires similar receptor conformations, whereas requirements for arrestin3 binding to M3R are distinct. Read full article

Nuclear magnetic resonance (NMR) spectroscopy allows the determination of atomic-level information on intermolecular interactions, molecular structure, and molecular dynamics in the cellular environment. This may be broadly divided into studies focused on obtaining detailed molecular information in the intracellular context ("in-cell") or those focused on characterizing molecules or events at the cell surface ("on-cell"). In this review, we outline some key NMR techniques applied for on-cell NMR studies through both solution- and solid-state NMR and survey studies that have used these techniques to uncover key information. In particular, we focus on the application of on-cell NMR spectroscopy to characterize ligand interactions with cell surface membrane proteins such as G-protein coupled receptors (GPCRs) and receptor tyrosine kinases. These techniques allow for quantification of binding affinities, competitive binding assays, delineation of ligands involved in binding, ligand bound-state conformational determination, evaluation of receptor structuring and dynamics, and inference of distance constraints characteristic of the ligand-receptor bound state. Interestingly, it is possible to avoid the barriers of production and purification of membrane proteins while obtaining directly physiologically relevant information through on-cell NMR. We also provide a brief survey of the applicability of on-cell NMR approaches to other classes of cell surface molecules. Read full article

Bitter taste receptors (TAS2Rs) are a poorly understood subgroup of G protein-coupled receptors (GPCRs). The experimental structure of these receptors has yet to be determined, and key-residues controlling their function remain mostly unknown. We designed an integrative approach to improve comparative modeling of TAS2Rs. Using current knowledge on class A GPCRs and existing experimental data in the literature as constraints, we pinpointed conserved motifs to entirely re-align the amino-acid sequences of TAS2Rs. We constructed accurate homology models of human TAS2Rs. As a test case, we examined the accuracy of the TAS2R16 model with site-directed mutagenesis and in vitro functional assays. This combination of in silico and in vitro results clarifies sequence-function relationships and proposes functional molecular switches that encode agonist sensing and downstream signaling mechanisms within mammalian TAS2Rs sequences. Read full article

Bioanalysis   2018 , 10  (19), 1609–1623. https://doi.org/10.4155/bio-2018-0150 . (11)      Showalter Bioconjugate Chem. 2013 , 24  (11), 1907–1916. https://doi.org/10.1021/bc400328m . (16)      Morrison Analytical Biochemistry   1988 , 174  (1), 101–120. https://doi.org/10.1016/0003-2697(88)90524-6 . (17 Principles and Biophysical Applications of Lanthanide-Based Probes. Nature Biotechnology   2003 , 21  (1), 86–89. https://doi.org/10.1038/nbt765 . (24)      Zwier, J.

Annual GPCRs-Targeted Drug Discovery Summit  is bringing together 80+ senior leaders across GPCR-focused biopharma NIS | Founding Co-Host NanoImaging Services (NIS) is a US-based structural biology partner offering a workflows, accelerate discovery, and drive breakthroughs in drug development, diagnostics, and disease biology Who should come This evening is open to pharma, biotech, academia, and students. This event is open to pharma, biotech, academia, and students.

Taken together, the lineup spans biophysics, kinetics, spatial biology, subcellular regulation, and program Sudarshan Rajagopal  — The Spatiotemporal Revolution in GPCR Biology. May 21. Dmitry Veprintsev  — Biophysical Approaches to Study Orphan GPCR Ligand Binding and Signalling. Alex Ball)  — A reagent gap that holds back the biology. Details coming soon.

on iPSC-derived translational models, and a new podcast episode with Joseph Kim on GPCR structural biology structural limitation when signaling outcomes depend on cell type, signaling complex assembly, and disease biology session examines how patient-derived induced pluripotent stem cells (iPSCs), organoid systems, and biosensor-based Joseph Kim: Structural Biology and Drug Discovery at GPCRs Cryo-EM has transformed how we visualize receptor–ligand This course examines prodrugs, biologics, irreversible inhibitors, and molecular glues — and how these

be interpreted when receptor behavior depends on cell type, signaling complex assembly, and disease biology This session with Terry Hébert will examine how iPSC-derived cardiomyocytes , organoid systems , and biosensor-based Organoid systems introduce multicellular organization, while biosensor-based approaches enable direct Biosensor-based assays enable direct measurement of signaling dynamics within complex biological systems GPCR brings together scientists working across GPCR biology, pharmacology, and drug discovery to examine

Separately, signaling outputs derived from heterologous systems often fail to reflect the biological environments they aim to represent. iPSC-derived cells, organoids, and biosensor-based assays introduce structural limitation when signaling outcomes depend on cell type, signaling complex assembly, and disease biology patient-derived induced pluripotent stem cells (iPSCs), iPSC-derived cardiomyocytes, organoid systems, and biosensor-based Key implications: Signaling outputs depend on biological context, not only receptor pharmacology iPSC

The data remains internally consistent—but the inferred mechanism diverges from the underlying biology Determines Pharmacology Allosteric behavior is not intrinsic to the ligand alone; it depends on the biological concepts laid out in the Corner will always be crucial for anyone studying or trying to modulate GPCR biology

What is needed is the biochemical resolution to tell them apart. starting from a synthetic ligand with measurable receptor activity and working backward to define receptor biology Given that the human genome encodes approximately 100 non-olfactory orphan GPCRs , the unexplored biology Even a well-characterized ligand in a cell-based assay faces the challenge of translation to complex biological in vitro  tools with phenotypic screening and whole-cell systems that more closely approximate human biology

The harder question is what those assay signals actually predict biologically. They fail because the interpretation of that data did not translate to biology outside the assay system Biology reveals targets. Even experienced biologists sometimes treat EC₅₀ as if it reflects ligand affinity. Pharmacologists interpret biological data.

Combining Two Technologies Into One to Measure A2A Fluorescent Competitive Binding Bioluminescence resonance proximity-based approach, a NanoLuc®luciferase   genetically fused  to the target protein   transfers  bioluminescence Design, Radiosynthesis, and Biodistribution of a New Potent and Selective Ligand for in Vivo Imaging Biochemical Pharmacology   1973 , 22  (23), 3099–3108. https://doi.org/10.1016/0006-2952(73)90196-2 .

It prevents overinterpretation of noise as biology.

16–18, 2026); Free fatty acid receptor 2 allosterism is defined by cellular context; Conformational biosensors These sessions span foundational pharmacology, receptor biology, modeling, translational strategy, and This partnership connects advanced assay and biology capabilities with the scientists and organizations Bitter taste receptors extend beyond taste biology. Job Listings  — GPCR-specific career opportunities across academia, biotech, and pharma.

Integrating Chemistry and Kinetics Early Biological activity alone does not define a viable series.

Volume II (Call for Papers, Deadline March 14) Signal transduction remains central to understanding GPCR biology A new volume dedicated to GPCR signal transduction invites contributions spanning cellular biochemistry Contribute to shaping scientific direction in cellular biochemistry.

Red-flagging toxicophores accelerates rational design cycles Computational prediction complements biological

GPCR scientists, translational pharmacologists, biotech discovery teams, and decision-makers who need

accelerates the design–test–learn cycle Drug discovery is faster when chemistry starts aligned with biology parenteral routes bypass absorption barriers, oral and topical delivery require navigating complex biological failures are rarely about route choice alone—they reflect mismatches between scaffold properties and biological

. 👉 In biotech, where cycles are long and feedback is slow, this gap becomes even more pronounced. Biotech teams operate across disciplines, timelines, and incentives. Attila runs focused strategy consultations for biotech founders who are ready to lead with clarity, not