Transmembrane a-helix 2 and 7 are important for small molecule-mediated activation of the GLP-1 receptor

Christina Rye Underwood; Sanne Møller Knudsen; Birgitte Schjellerup Wulff; Hans Bräuner-Osborne; Jesper Lau; Lotte Bjerre Knudsen; Günther H.J. Peters; Steffen Reedtz-Runge

doi:10.1159/000334338

Transmembrane a-helix 2 and 7 are important for small molecule-mediated activation of the GLP-1 receptor

Christina Rye Underwood, Sanne Møller Knudsen, Birgitte Schjellerup Wulff, Hans Bräuner-Osborne, Jesper Lau, Lotte Bjerre Knudsen, Günther H.J. Peters, Steffen Reedtz-Runge

7 Citations (Scopus)

Abstract

Glucagon-like peptide-1 (GLP-1) activates the GLP-1 receptor (GLP-1R), which belongs to family B of the G-protein-coupled receptors. We previously identified a selective small molecule ligand, compound 2, that acted as a full agonist and allosteric modulator of GLP-1R. In this study, the structurally related small molecule, compound 3, stimulated cAMP production from GLP-1R, but not from the homologous glucagon receptor (GluR). The receptor selectivity encouraged a chimeric receptor approach to identify domains important for compound 3-mediated activation of GLP-1R. A subsegment of the GLP-1R transmembrane domain containing TM2 to TM5 was sufficient to transfer compound 3 responsiveness to GluR. Therefore, divergent residues in this subsegment of GLP-1R and GluR are responsible for the receptor selectivity of compound 3. Functional analyses of other chimeric receptors suggested that the existence of a helix-helix interface between TM1 and TM7 is important for the compound 3 response. Furthermore, site-directed mutagenesis revealed that a Phe195-Leu substitution in TM2 and a Thr391-Ala substitution in TM7 increased and decreased the efficacy of compound 3 without disturbing the potency or efficacy of GLP-1. Collectively, differential effects of receptor mutations suggest that TM2 and/or TM7 are important for compound 3-mediated activation of GLP-1R.

Original language	English
Journal	Pharmacology
Volume	88
Issue number	5-6
Pages (from-to)	340-348
Number of pages	9
ISSN	0031-7012
DOIs	https://doi.org/10.1159/000334338
Publication status	Published - Dec 2011

Keywords

Cyclic AMP
Glucagon
Glucagon-Like Peptide 1
HEK293 Cells
Humans
Ligands
Models, Molecular
Peptide Fragments
Protein Structure, Secondary
Receptors, Glucagon

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@article{3fb5186da4284eb9834a6dc447b15413,

title = "Transmembrane a-helix 2 and 7 are important for small molecule-mediated activation of the GLP-1 receptor",

abstract = "Glucagon-like peptide-1 (GLP-1) activates the GLP-1 receptor (GLP-1R), which belongs to family B of the G-protein-coupled receptors. We previously identified a selective small molecule ligand, compound 2, that acted as a full agonist and allosteric modulator of GLP-1R. In this study, the structurally related small molecule, compound 3, stimulated cAMP production from GLP-1R, but not from the homologous glucagon receptor (GluR). The receptor selectivity encouraged a chimeric receptor approach to identify domains important for compound 3-mediated activation of GLP-1R. A subsegment of the GLP-1R transmembrane domain containing TM2 to TM5 was sufficient to transfer compound 3 responsiveness to GluR. Therefore, divergent residues in this subsegment of GLP-1R and GluR are responsible for the receptor selectivity of compound 3. Functional analyses of other chimeric receptors suggested that the existence of a helix-helix interface between TM1 and TM7 is important for the compound 3 response. Furthermore, site-directed mutagenesis revealed that a Phe195-Leu substitution in TM2 and a Thr391-Ala substitution in TM7 increased and decreased the efficacy of compound 3 without disturbing the potency or efficacy of GLP-1. Collectively, differential effects of receptor mutations suggest that TM2 and/or TM7 are important for compound 3-mediated activation of GLP-1R.",

keywords = "Cyclic AMP, Glucagon, Glucagon-Like Peptide 1, HEK293 Cells, Humans, Ligands, Models, Molecular, Peptide Fragments, Protein Structure, Secondary, Receptors, Glucagon",

author = "Underwood, {Christina Rye} and Knudsen, {Sanne M{\o}ller} and Wulff, {Birgitte Schjellerup} and Hans Br{\"a}uner-Osborne and Jesper Lau and Knudsen, {Lotte Bjerre} and Peters, {G{\"u}nther H.J.} and Steffen Reedtz-Runge",

note = "Keywords: G-protein coupled receptor, small molecule agonist, cAMP, mutagenesis ",

year = "2011",

month = dec,

doi = "10.1159/000334338",

language = "English",

volume = "88",

pages = "340--348",

journal = "Pharmacology",

issn = "0031-7012",

publisher = "S Karger AG",

number = "5-6",

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TY - JOUR

T1 - Transmembrane a-helix 2 and 7 are important for small molecule-mediated activation of the GLP-1 receptor

AU - Underwood, Christina Rye

AU - Knudsen, Sanne Møller

AU - Wulff, Birgitte Schjellerup

AU - Bräuner-Osborne, Hans

AU - Lau, Jesper

AU - Knudsen, Lotte Bjerre

AU - Peters, Günther H.J.

AU - Reedtz-Runge, Steffen

N1 - Keywords: G-protein coupled receptor, small molecule agonist, cAMP, mutagenesis

PY - 2011/12

Y1 - 2011/12

N2 - Glucagon-like peptide-1 (GLP-1) activates the GLP-1 receptor (GLP-1R), which belongs to family B of the G-protein-coupled receptors. We previously identified a selective small molecule ligand, compound 2, that acted as a full agonist and allosteric modulator of GLP-1R. In this study, the structurally related small molecule, compound 3, stimulated cAMP production from GLP-1R, but not from the homologous glucagon receptor (GluR). The receptor selectivity encouraged a chimeric receptor approach to identify domains important for compound 3-mediated activation of GLP-1R. A subsegment of the GLP-1R transmembrane domain containing TM2 to TM5 was sufficient to transfer compound 3 responsiveness to GluR. Therefore, divergent residues in this subsegment of GLP-1R and GluR are responsible for the receptor selectivity of compound 3. Functional analyses of other chimeric receptors suggested that the existence of a helix-helix interface between TM1 and TM7 is important for the compound 3 response. Furthermore, site-directed mutagenesis revealed that a Phe195-Leu substitution in TM2 and a Thr391-Ala substitution in TM7 increased and decreased the efficacy of compound 3 without disturbing the potency or efficacy of GLP-1. Collectively, differential effects of receptor mutations suggest that TM2 and/or TM7 are important for compound 3-mediated activation of GLP-1R.

AB - Glucagon-like peptide-1 (GLP-1) activates the GLP-1 receptor (GLP-1R), which belongs to family B of the G-protein-coupled receptors. We previously identified a selective small molecule ligand, compound 2, that acted as a full agonist and allosteric modulator of GLP-1R. In this study, the structurally related small molecule, compound 3, stimulated cAMP production from GLP-1R, but not from the homologous glucagon receptor (GluR). The receptor selectivity encouraged a chimeric receptor approach to identify domains important for compound 3-mediated activation of GLP-1R. A subsegment of the GLP-1R transmembrane domain containing TM2 to TM5 was sufficient to transfer compound 3 responsiveness to GluR. Therefore, divergent residues in this subsegment of GLP-1R and GluR are responsible for the receptor selectivity of compound 3. Functional analyses of other chimeric receptors suggested that the existence of a helix-helix interface between TM1 and TM7 is important for the compound 3 response. Furthermore, site-directed mutagenesis revealed that a Phe195-Leu substitution in TM2 and a Thr391-Ala substitution in TM7 increased and decreased the efficacy of compound 3 without disturbing the potency or efficacy of GLP-1. Collectively, differential effects of receptor mutations suggest that TM2 and/or TM7 are important for compound 3-mediated activation of GLP-1R.

KW - Cyclic AMP

KW - Glucagon

KW - Glucagon-Like Peptide 1

KW - HEK293 Cells

KW - Humans

KW - Ligands

KW - Models, Molecular

KW - Peptide Fragments

KW - Protein Structure, Secondary

KW - Receptors, Glucagon

U2 - 10.1159/000334338

DO - 10.1159/000334338

M3 - Journal article

C2 - 22134089

SN - 0031-7012

VL - 88

SP - 340

EP - 348

JO - Pharmacology

JF - Pharmacology

IS - 5-6

ER -

Transmembrane a-helix 2 and 7 are important for small molecule-mediated activation of the GLP-1 receptor

Abstract

Keywords

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