GPCR NaVa database: natural variants in human G protein-coupled receptors

Kazius, Jeroen; Wurdinger, Kerstin; Iterson, Maarten van; Kok, Joost N.; Bäck, Thomas; IJzerman, Adriaan P.

doi:10.1002/humu.20638

Cited by 47 publications

(32 citation statements)

References 26 publications

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“…1, A and B, respectively). Each incretin receptor variant appeared in the NaVa (Natural Variants) database that catalogs known human GPCR polymorphisms (frequency Ͼ1%), as well as rarer mutations (Kazius et al, 2008). As outlined in the discussion, three GIP-R variants have been described previously in the literature (Kubota et al, 1996;Almind et al, 1998).…”

Section: Resultsmentioning

confidence: 99%

Pharmacological Characterization of Human Incretin Receptor Missense Variants

Fortin¹,

Schroeder²,

Zhu³

et al. 2009

J Pharmacol Exp Ther

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Glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) are gut-derived incretin hormones that regulate blood glucose levels. In addition to their widely accepted insulinotropic role, there is evidence that GLP-1 modulates feeding behavior and GIP regulates lipid metabolism, thereby promoting postprandial fat deposition. In this study, we investigated whether naturally occurring polymorphisms in the GLP-1 receptor (GLP-1R) and the GIP receptor (GIP-R) affect the pharmacological properties of these proteins. After transient expression of the receptors in human embryonic kidney 293 cells, basal and ligand-induced cAMP production were assessed by use of luciferase reporter gene assays. Our data reveal that the wild-type GIP-R displays a considerable degree of ligand-independent activity. In comparison, the GIP-R variants C46S, G198C, R316L, and E354Q show a marked decrease in basal signaling that may, at least in part, be explained by reduced cell surface expression. When stimulated with GIP, the C46S and R316L mutants display significantly reduced potency (Ͼ1000 and 25-fold, respectively) compared with wild type. Complementary competition binding assays further demonstrate that the C46S variant fails to bind radio-iodinated GIP, whereas all other GIP-R mutants maintain normal ligand affinity. In contrast to the GIP-R, the wild-type GLP-1R lacks constitutive activity. Furthermore, none of the 10 GLP-1R missense mutations showed an alteration in pharmacological properties versus wild type. The extent to which abnormalities in GIP-R function may lead to physiological changes or affect drug sensitivity in selected populations (e.g., obese, diabetic individuals) remains to be further investigated.The incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) are homologous peptides released from intestinal enteroendocrine cells in response to food intake. Both hormones are important modulators of metabolic function. In the pancreas, GLP-1 and GIP potentiate nutrient-stimulated insulin secretion and promote the expansion of pancreatic islet mass via induction of ␤-cell proliferation and survival (Kim et al., 2005;Kim and Egan, 2008).

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Section: Resultsmentioning

confidence: 99%

Pharmacological Characterization of Human Incretin Receptor Missense Variants

Fortin¹,

Schroeder²,

Zhu³

et al. 2009

J Pharmacol Exp Ther

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“…Thirteen nonsynonymous single amino acid substitutions in the human MOR (A6V, N40D, D51N, G63V, S66F, S147C, N190K, R260H, R265C, R265H, S268P, D274N, and V293I) were selected for analysis based on a review of the NaVa database (Kazius et al, 2008) and of the literature (Befort et al, 2001;Wang et al, 2001;Lotsch and Geisslinger, 2005). The position of each amino acid substitution is illustrated in a diagram of the MOR (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In the current study, 13 nonsynonymous single amino acid changes in the MOR were selected for pharmacological analysis from the NaVa (Natural Variants) database, which catalogs known human GPCR polymorphisms (frequency Ͼ1%) as well as rarer mutations (Kazius et al, 2008). Our investigations suggest that selected MOR variants show reduced or absent agonist function.…”

Section: Introductionmentioning

confidence: 99%

The μ-Opioid Receptor Variant N190K Is Unresponsive to Peptide Agonists yet Can be Rescued by Small-Molecule Drugs

Fortin

Ci²,

Schroeder³

et al. 2010

Mol Pharmacol

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The -opioid receptor (MOR) plays an important role in modulating analgesia, feeding behavior, and a range of autonomic functions. In the current study, we investigated the degree to which 13 naturally occurring missense mutations affect the pharmacological properties of the human MOR. After expression of each receptor in human embryonic kidney 293 cells, signaling (G␣ i/o -mediated) induced by peptide agonists was assessed using luciferase reporter gene assays. Multiple mutants (S66F, S147C, R260H, R265C, R265H, and S268P) show a significant reduction in agonist potency. At the N190K variant, agonist-mediated signaling was essentially absent. Enzymelinked immunosorbent assay, microscopic analysis, and radioligand binding assays revealed that this mutant shows markedly reduced cell-surface expression, whereas all other receptor variants were expressed at normal levels. Surface expression of the N190K variant could be increased by incubation with the alkaloid agonist buprenorphine or with either naltrexone or naloxone, structurally related MOR antagonists. We were surprised to find that both putative antagonists, despite being inactive at the wild-type MOR, triggered a concentration-dependent increase in N190K receptor-mediated signaling. In contrast, peptidic ligands failed to promote expression or rescue function of the N190K mutant. Subsequent analysis of the N190K variant in an ethnically diverse cohort identified this isoform in a subgroup of African Americans. Taken together, our studies reveal that the N190K mutation leads to severe functional alterations and, in parallel, changes the response to established MOR ligands. The extent to which this mutation results in physiological abnormalities or affects drug sensitivity in selected populations (e.g., those with chronic pain or addiction) remains to be investigated.

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“…Other recent integration-promoting databases related to GPCRs are RINGdb [92] and the GPCR-NaVa (GPCR Natural Variants) database [93]. RINGdb is a relational database of GPCRs and the regulators of G-protein signaling (RGS), a family of proteins that share a conserved signature domain (RGS domain) that binds directly to activated G subunits to modulate G-protein signaling.…”

Section: Dedicated Databases and Serversmentioning

confidence: 99%

Advances in the development and application of computational methodologies for structural modeling of G-protein-coupled receptors

Mobarec¹,

Filizola²

2008

Expert Opinion on Drug Discovery

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Background-Despite the large amount of experimental data accumulated in the past decade on G-protein coupled receptor (GPCR) structure and function, understanding of the molecular mechanisms underlying GPCR signaling is still far from being complete, thus impairing the design of effective and selective pharmaceuticals.Objective-Understanding of GPCR function has been challenged even further by more recent experimental evidence that several of these receptors are organized in the cell membrane as homoor hetero-oligomers, and that they may exhibit unique pharmacological properties. Given the complexity of these new signaling systems, researcher's efforts are turning increasingly to molecular modeling, bioinformatics and computational simulations for mechanistic insights of GPCR functional plasticity.Methods-We review here current advances in the development and application of computational approaches to improve prediction of GPCR structure and dynamics, thus enhancing current understanding of GPCR signaling.Results/Conclusions-Models resulting from use of these computational approaches further supported by experiments are expected to help elucidate the complex allosterism that propagates through GPCR complexes, ultimately aiming at successful structure-based rational drug design.

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GPCR NaVa database: natural variants in human G protein-coupled receptors

Cited by 47 publications

References 26 publications

Pharmacological Characterization of Human Incretin Receptor Missense Variants

Pharmacological Characterization of Human Incretin Receptor Missense Variants

The μ-Opioid Receptor Variant N190K Is Unresponsive to Peptide Agonists yet Can be Rescued by Small-Molecule Drugs

Advances in the development and application of computational methodologies for structural modeling of G-protein-coupled receptors

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