A5003364568- Receptor Mechanisms and Signaling
- Neuropeptides and Animal Physiology
- Pharmacological Effects and Assays
- Neuroscience and Neuropharmacology Research
- Biochemical Analysis and Sensing Techniques
- Mass Spectrometry Techniques and Applications
- Drug Transport and Resistance Mechanisms
- Adenosine and Purinergic Signaling
- Protein Kinase Regulation and GTPase Signaling
- Asthma and respiratory diseases
- Pancreatic function and diabetes
- Diabetes Treatment and Management
- Crystallization and Solubility Studies
- Chemokine receptors and signaling
- Monoclonal and Polyclonal Antibodies Research
- Regulation of Appetite and Obesity
- Bacterial Genetics and Biotechnology
- X-ray Diffraction in Crystallography
- RNA and protein synthesis mechanisms
- Helicobacter pylori-related gastroenterology studies
- Peroxisome Proliferator-Activated Receptors
- Microbial Metabolic Engineering and Bioproduction
- Sphingolipid Metabolism and Signaling
- Phosphodiesterase function and regulation
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
University of Glasgow
2015-2024
Jena University Hospital
2023
Institute of Molecular and Cell Biology
2017
University of Southern Denmark
2014
Universitat Autònoma de Barcelona
2013
Novartis (United Kingdom)
2013
LifeArc
2012
University of California, Irvine
1987
Stanford University
1987
Abstract Despite the discovery of heterotrimeric αβγ G proteins ∼25 years ago, their selective perturbation by cell-permeable inhibitors remains a fundamental challenge. Here we report that plant-derived depsipeptide FR900359 (FR) is ideally suited to this task. Using multifaceted approach systematically characterize FR as inhibitor Gq/11/14 over all other mammalian Gα isoforms and elaborate its molecular mechanism action. We also use investigate whether inhibition Gq an effective...
Various foods are associated with effects against metabolic diseases such as insulin resistance and type 2 diabetes; however, their mechanisms of action mostly unclear. Fatty acids may contribute by acting precursors signalling molecules or direct activity on receptors. The medium- long-chain NEFA receptor FFA1 (free fatty acid 1, previously known GPR40) has been linked to enhancement glucose-stimulated secretion, whereas FFA4 4, GPR120) insulin-sensitising anti-inflammatory effects, both...
Free fatty acid receptors 1 to 4 (FFA1 FFA4) are class A G protein–coupled (GPCRs). FFA1 FFA3 share substantial sequence similarity, whereas FFA4 is unrelated. However, and activated by long-chain acids, while FFA2 respond short-chain acids generated intestinal microbiota. FFA1, FFA2, potential drug targets for metabolic inflammatory conditions. Here, we determined the active structures of bound docosahexaenoic acid, synthetic agonist TUG-891, butyrate-bound each complexed with an engineered...
The structural organization and regulation of the genes involved in short-chain fatty acid degradation Escherichia coli, referred to as ato system, have been studied by a combination classic genetic recombinant DNA techniques. A plasmid containing 6.2-kilobase region E. coli chromosome was able complement mutations genes, atoA (acetyl-coenzyme [CoA]:acetoacetyl [AA]-CoA transferase) atoB (thiolase II), well regulatory locus, atoC. Complementation studies performed with mutants defective...
FFA2 and FFA3 are closely related G protein-coupled receptors that bind respond to short chain fatty acids. (FFA2 the provisional International Union of Pharmacology designations for previously called GPR43 GPR41, respectively.) Sequence comparisons between these two alignments with receptor FFA1, linked homology modeling based on atomic level structure bovine rhodopsin, indicated potential polar residues within transmembrane helix bundle play important roles in ligand recognition function....
The poorly characterized G-protein-coupled receptor GPR35 has been suggested as a potential exploratory target for the treatment of both metabolic disorders and hypertension. It also indicated to play an important role in immune modulation. A major impediment validation these concepts further study this paucity pharmacological tools that interact with GPR35. Using receptor-β-arrestin-2 interaction assay human rat orthologues GPR35, we identified number compounds possessing agonist activity....
Volatile small molecules, including the short-chain fatty acids (SCFAs), acetate and propionate, released by gut microbiota from catabolism of nondigestible starches, can act in a hormone-like fashion via specific G-protein-coupled receptors (GPCRs). The primary GPCR targets for these SCFAs are FFA2 FFA3. Using transgenic mice which was replaced an altered form called Designer Receptor Exclusively Activated Drugs (FFA2-DREADD), but FFA3 is unaltered, newly identified FFA2-DREADD agonist...
Abstract GPR84 is a unique orphan G protein-coupled receptor (GPCR) that can be activated by endogenous medium-chain fatty acids (MCFAs). The signaling of largely pro-inflammatory, which augment inflammatory response, and also functions as pro-phagocytic to enhance phagocytic activities macrophages. In this study, we show the activation synthetic agonist 6-OAU synergize with blockade CD47 on cancer cells induce phagocytosis We determine high-resolution structure GPR84-G i complex 6-OAU. This...
Coexpression of the MAS proto-oncogene with angiotensin II type 1 (AT(1)) receptor in CHO-K1 cells has been reported to increase number [(3)H]angiotensin II-binding sites, although does not bind II. In HEK293 stably expressing AT(1) receptor-cyan fluorescent protein (CFP), MAS-yellow (YFP) expression from an inducible locus caused strong up-regulation receptor-CFP amounts and binding levels. The time course was also markedly slower than that induction expression. These effects were mimicked...
The endogenous ligands for free fatty acid receptor 1 (FFA1) are medium and longer chain acids. However, a range of selective, small molecule have recently been developed as tool compounds to explore the therapeutic potential this receptor, whereas clinically employed thiazolidinedione "glitazone" drugs also agonists at FFA1. Each these classes agonist was able promote phosphorylation ERK1/2 mitogen-activated protein (MAP) kinases in cells express human FFA1 on demand. although both lauric...
GPR35 is a poorly characterized G protein-coupled receptor at which kynurenic acid has been suggested to be the endogenous ligand. We wished test this and develop assays appropriate for study of receptor.Human rat orthologues were engineered expressed developed assess interaction with β-arrestin-2, activation Gα₁₃ agonist-induced internalization.GPR35-β-arrestin-2 confirmed that both tryptophan metabolite synthetic ligand zaprinast had agonist action each orthologue. Zaprinast was...
Free fatty acid receptor 1 (FFA1 or GPR40) enhances glucose-stimulated insulin secretion from pancreatic β-cells and currently attracts high interest as a new target for the treatment of type 2 diabetes. We here report discovery highly potent FFA1 agonist with favorable physicochemical pharmacokinetic properties. The compound efficiently normalizes glucose tolerance in diet-induced obese mice, an effect that is fully sustained after 29 days chronic dosing.
Analysis of the roles short chain fatty acid receptor, free 3 receptor (FFA3), has been severely limited by low potency its endogenous ligands, crossover function these on closely related 2 and a dearth FFA3-selective synthetic ligands. From series hexahydroquinolone-3-carboxamides, we demonstrate that 4-(furan-2-yl)-2-methyl-5-oxo-<i>N</i>-(<i>o</i>-tolyl)-1,4,5,6,7,8-hexahydroquinoline-3-carboxamide is selective moderately potent positive allosteric modular (PAM)-agonist FFA3 receptor....
Lack of high potency agonists has restricted analysis the G protein–coupled receptor GPR35. Moreover, marked variation in and/or affinity current ligands between human and rodent orthologs GPR35 limited their productive use models physiology. Based on reported modest antiasthma antiallergic cromolyn disodium nedocromil sodium, we identified related compounds lodoxamide bufrolin as Unlike previously that are highly selective for GPR35, both displayed equivalent at rat Further synthetic...
Abstract G protein-coupled receptor (GPR)35 is highly expressed in the gastro-intestinal tract, predominantly colon epithelial cells (CEC), and has been associated with inflammatory bowel diseases (IBD), suggesting a role gastrointestinal inflammation. The enterotoxigenic Bacteroides fragilis (ETBF) toxin (BFT) an important virulence factor causing gut inflammation humans animal models. We identified that BFT signals through GPR35. Blocking GPR35 function CECs using antagonist ML145,...
G protein-coupled receptor 35 (GPR35) is poorly characterized but nevertheless has been revealed to have diverse roles in areas including lower gut inflammation and pain. The development of novel reagents tools will greatly enhance analysis GPR35 functions health disease. Here, we used mass spectrometry, mutagenesis, [32P] orthophosphate labeling identify that all five hydroxy-amino acids the C-terminal tail human GPR35a became phosphorylated response agonist occupancy that, apart from...
Variation in pharmacology and function of ligands at species orthologs can be a confounding feature understanding the biology role poorly characterized receptors. Substantial selectivity potency number GPR35 agonists has previously been demonstrated between human rat this G protein-coupled receptor. Via bioluminescence resonance energy transfer-based assay induced interactions β-arrestin-2, addition mouse ortholog to such studies indicated that, as for ortholog, murine displayed very low...
Medium chain fatty acids can activate the pro-inflammatory receptor GPR84 but so also molecules related to 3,3'-diindolylmethane. 3,3'-Diindolylmethane and decanoic acid acted as strong positive allosteric modulators of function each other analysis showed affinity 3,3'-diindolylmethane be at least 100 fold higher. Methyl decanoate was not an agonist GPR84. This implies a key role in binding for carboxylic acid. Via homology modelling we predicted confirmed integral arginine172, located 2nd...
Abstract The chemokine CXCL17 is associated with the innate response in mucosal tissues but poorly characterized. Similarly, G protein–coupled receptor GPR35, expressed by monocytes and mast cells, has been implicated immune response, although its precise role ill-defined. A recent manuscript reported that GPR35 was able to signal CXCL17, which we set out confirm this study. readily using transfection systems failed assays of β-arrestin recruitment, inositol phosphate production, calcium...
GPR84 is a medium chain free fatty acid-binding G-protein-coupled receptor associated with inflammatory and fibrotic diseases. As the only reported antagonist of (PBI-4050) that displays relatively low potency selectivity, clear need exists for an improved modulator. Structural optimization hit 1, identified through high-throughput screening, led to identification potent selective inhibitor GLPG1205 (36). Compared initial hit, 36 showed in guanosine 5′-O-[γ-thio]triphosphate assay, exhibited...
GPR84 is an immune cell-expressed, proinflammatory receptor currently being assessed as a therapeutic target in conditions including fibrosis and inflammatory bowel disease. Although it was previously shown that the orthosteric activators 2-HTP 6-OAU promoted its interactions with arrestin-3, G protein-biased agonist DL-175 did not. Here, we show replacement of all 21 serine threonine residues within i-loop 3 GPR84, but not two serines C-terminal tail, eliminated incorporation [32P] greatly...
The expression of the Ato enzymes, acetyl coenzyme A:acetoacetyl A transferase and thiolase II, is required for growth Escherichia coli on short-chain fatty acids. structural genes these atoD, atoA, atoB, respectively, make up ato operon. 48-kilodalton protein encoded by atoC was synthesis or activation enzymes. enzyme activities inducible in atoC+ strains, constitutive atoCc noninducible mutants. Merodiploid studies demonstrated that allele trans-dominant to allele. To study action...
Despite recent advances in structural definition of GPCR-G protein complexes, the basis receptor selectivity between G proteins remains unclear. The Gα12 and Gα13 subtypes together form least studied group heterotrimeric proteins. protein-coupled 35 (GPR35) has been suggested to couple efficiently but weakly Gα12. Using combinations cells genome-edited not express bioluminescence resonance energy transfer-based sensors, we confirmed marked GPR35 for Gα13. Incorporating Gα12/Gα13 chimeras...