Evidence that interaction between conserved residues in transmembrane helices 2, 3, and 7 are crucial for human VPAC1 receptor activation

Anton O Chugunov, John Simms, David R Poyner, Yves Dehouck, Marianne Rooman, Dimitri Gilis, Ingrid Langer

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The VPAC(1) receptor belongs to family B of G protein-coupled receptors (GPCR-B) and is activated upon binding of the vasoactive intestinal peptide (VIP). Despite the recent determination of the structure of the N terminus of several members of this receptor family, little is known about the structure of the transmembrane (TM) region and about the molecular mechanisms leading to activation. In the present study, we designed a new structural model of the TM domain and combined it with experimental mutagenesis experiments to investigate the interaction network that governs ligand binding and receptor activation. Our results suggest that this network involves the cluster of residues Arg(188) in TM2, Gln(380) in TM7, and Asn(229) in TM3. This cluster is expected to be altered upon VIP binding, because Arg(188) has been shown previously to interact with Asp(3) of VIP. Several point mutations at positions 188, 229, and 380 were experimentally characterized and were shown to severely affect VIP binding and/or VIP-mediated cAMP production. Double mutants built from reciprocal residue exchanges exhibit strong cooperative or anticooperative effects, thereby indicating the spatial proximity of residues Arg(188), Gln(380), and Asn(229). Because these residues are highly conserved in the GPCR-B family, they can moreover be expected to have a general role in mediating function.

Original languageEnglish
Pages (from-to)394-401
Number of pages8
JournalMolecular Pharmacology
Volume78
Issue number3
DOIs
Publication statusPublished - Sep 2010

Fingerprint

Receptors, Vasoactive Intestinal Polypeptide, Type I
Vasoactive Intestinal Peptide
Structural Models
Viperidae
G-Protein-Coupled Receptors
Point Mutation
Mutagenesis
Ligands

Keywords

  • Animals
  • Asparagine
  • Cellular Structures
  • Cricetinae
  • Humans
  • Mutagenesis
  • Protein Structure, Secondary
  • Receptors, G-Protein-Coupled
  • Receptors, Vasoactive Intestinal Polypeptide, Type I
  • Vasoactive Intestinal Peptide
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Evidence that interaction between conserved residues in transmembrane helices 2, 3, and 7 are crucial for human VPAC1 receptor activation. / Chugunov, Anton O; Simms, John; Poyner, David R; Dehouck, Yves; Rooman, Marianne; Gilis, Dimitri; Langer, Ingrid.

In: Molecular Pharmacology, Vol. 78, No. 3, 09.2010, p. 394-401.

Research output: Contribution to journalArticle

Chugunov, Anton O ; Simms, John ; Poyner, David R ; Dehouck, Yves ; Rooman, Marianne ; Gilis, Dimitri ; Langer, Ingrid. / Evidence that interaction between conserved residues in transmembrane helices 2, 3, and 7 are crucial for human VPAC1 receptor activation. In: Molecular Pharmacology. 2010 ; Vol. 78, No. 3. pp. 394-401.
@article{9ee229233fd841dd932545806f416757,
title = "Evidence that interaction between conserved residues in transmembrane helices 2, 3, and 7 are crucial for human VPAC1 receptor activation",
abstract = "The VPAC(1) receptor belongs to family B of G protein-coupled receptors (GPCR-B) and is activated upon binding of the vasoactive intestinal peptide (VIP). Despite the recent determination of the structure of the N terminus of several members of this receptor family, little is known about the structure of the transmembrane (TM) region and about the molecular mechanisms leading to activation. In the present study, we designed a new structural model of the TM domain and combined it with experimental mutagenesis experiments to investigate the interaction network that governs ligand binding and receptor activation. Our results suggest that this network involves the cluster of residues Arg(188) in TM2, Gln(380) in TM7, and Asn(229) in TM3. This cluster is expected to be altered upon VIP binding, because Arg(188) has been shown previously to interact with Asp(3) of VIP. Several point mutations at positions 188, 229, and 380 were experimentally characterized and were shown to severely affect VIP binding and/or VIP-mediated cAMP production. Double mutants built from reciprocal residue exchanges exhibit strong cooperative or anticooperative effects, thereby indicating the spatial proximity of residues Arg(188), Gln(380), and Asn(229). Because these residues are highly conserved in the GPCR-B family, they can moreover be expected to have a general role in mediating function.",
keywords = "Animals, Asparagine, Cellular Structures, Cricetinae, Humans, Mutagenesis, Protein Structure, Secondary, Receptors, G-Protein-Coupled, Receptors, Vasoactive Intestinal Polypeptide, Type I, Vasoactive Intestinal Peptide, Journal Article, Research Support, Non-U.S. Gov't",
author = "Chugunov, {Anton O} and John Simms and Poyner, {David R} and Yves Dehouck and Marianne Rooman and Dimitri Gilis and Ingrid Langer",
year = "2010",
month = "9",
doi = "10.1124/mol.110.063578",
language = "English",
volume = "78",
pages = "394--401",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "3",

}

TY - JOUR

T1 - Evidence that interaction between conserved residues in transmembrane helices 2, 3, and 7 are crucial for human VPAC1 receptor activation

AU - Chugunov, Anton O

AU - Simms, John

AU - Poyner, David R

AU - Dehouck, Yves

AU - Rooman, Marianne

AU - Gilis, Dimitri

AU - Langer, Ingrid

PY - 2010/9

Y1 - 2010/9

N2 - The VPAC(1) receptor belongs to family B of G protein-coupled receptors (GPCR-B) and is activated upon binding of the vasoactive intestinal peptide (VIP). Despite the recent determination of the structure of the N terminus of several members of this receptor family, little is known about the structure of the transmembrane (TM) region and about the molecular mechanisms leading to activation. In the present study, we designed a new structural model of the TM domain and combined it with experimental mutagenesis experiments to investigate the interaction network that governs ligand binding and receptor activation. Our results suggest that this network involves the cluster of residues Arg(188) in TM2, Gln(380) in TM7, and Asn(229) in TM3. This cluster is expected to be altered upon VIP binding, because Arg(188) has been shown previously to interact with Asp(3) of VIP. Several point mutations at positions 188, 229, and 380 were experimentally characterized and were shown to severely affect VIP binding and/or VIP-mediated cAMP production. Double mutants built from reciprocal residue exchanges exhibit strong cooperative or anticooperative effects, thereby indicating the spatial proximity of residues Arg(188), Gln(380), and Asn(229). Because these residues are highly conserved in the GPCR-B family, they can moreover be expected to have a general role in mediating function.

AB - The VPAC(1) receptor belongs to family B of G protein-coupled receptors (GPCR-B) and is activated upon binding of the vasoactive intestinal peptide (VIP). Despite the recent determination of the structure of the N terminus of several members of this receptor family, little is known about the structure of the transmembrane (TM) region and about the molecular mechanisms leading to activation. In the present study, we designed a new structural model of the TM domain and combined it with experimental mutagenesis experiments to investigate the interaction network that governs ligand binding and receptor activation. Our results suggest that this network involves the cluster of residues Arg(188) in TM2, Gln(380) in TM7, and Asn(229) in TM3. This cluster is expected to be altered upon VIP binding, because Arg(188) has been shown previously to interact with Asp(3) of VIP. Several point mutations at positions 188, 229, and 380 were experimentally characterized and were shown to severely affect VIP binding and/or VIP-mediated cAMP production. Double mutants built from reciprocal residue exchanges exhibit strong cooperative or anticooperative effects, thereby indicating the spatial proximity of residues Arg(188), Gln(380), and Asn(229). Because these residues are highly conserved in the GPCR-B family, they can moreover be expected to have a general role in mediating function.

KW - Animals

KW - Asparagine

KW - Cellular Structures

KW - Cricetinae

KW - Humans

KW - Mutagenesis

KW - Protein Structure, Secondary

KW - Receptors, G-Protein-Coupled

KW - Receptors, Vasoactive Intestinal Polypeptide, Type I

KW - Vasoactive Intestinal Peptide

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1124/mol.110.063578

DO - 10.1124/mol.110.063578

M3 - Article

VL - 78

SP - 394

EP - 401

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 3

ER -