Functionally Different Agonists Induce Distinct Conformations in the G Protein Coupling Domain of the β2 Adrenergic Receptor

Pejman Ghanouni, Zygmunt Gryczynski, Jacqueline J. Steenhuis, Tae Weon Lee, David L. Farrens, Joseph R. Lakowicz, Brian K. Kobilka

Research output: Contribution to journalArticle

318 Citations (Scopus)

Abstract

G protein-coupled receptors represent the largest class of drug discovery targets. Drugs that activate G protein-coupled receptors are classified as either agonists or partial agonists. To study the mechanism whereby these different classes of activating ligands modulate receptor function, we directly monitored ligand-induced conformational changes in the G protein-coupling domain of the β2 adrenergic receptor. Fluorescence lifetime analysis of a reporter fluorophore covalently attached to this domain revealed that, in the absence of ligands, this domain oscillates around a single detectable conformation. Binding to an antagonist does not change this conformation but does reduce the flexibility of the domain. However, when the β2 adrenergic receptor is bound to a full agonist, the G protein coupling domain exists in two distinct conformations. Moreover, the conformations induced by a full agonist can be distinguished from those induced by partial agonists. These results provide new insight into the structural consequence of antagonist binding and the basis of agonism and partial agonism.

Original languageEnglish
Pages (from-to)24433-24436
Number of pages4
JournalJournal of Biological Chemistry
Volume276
Issue number27
DOIs
StatePublished - 6 Jul 2001

Fingerprint

GTP-Binding Proteins
Adrenergic Receptors
Conformations
G-Protein-Coupled Receptors
Ligands
Drug Discovery
Fluorophores
Fluorescence
Pharmaceutical Preparations
Protein Domains

Cite this

Ghanouni, Pejman ; Gryczynski, Zygmunt ; Steenhuis, Jacqueline J. ; Lee, Tae Weon ; Farrens, David L. ; Lakowicz, Joseph R. ; Kobilka, Brian K. / Functionally Different Agonists Induce Distinct Conformations in the G Protein Coupling Domain of the β2 Adrenergic Receptor. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 27. pp. 24433-24436.
@article{3675835c38884576adc2f7d43179e5e4,
title = "Functionally Different Agonists Induce Distinct Conformations in the G Protein Coupling Domain of the β2 Adrenergic Receptor",
abstract = "G protein-coupled receptors represent the largest class of drug discovery targets. Drugs that activate G protein-coupled receptors are classified as either agonists or partial agonists. To study the mechanism whereby these different classes of activating ligands modulate receptor function, we directly monitored ligand-induced conformational changes in the G protein-coupling domain of the β2 adrenergic receptor. Fluorescence lifetime analysis of a reporter fluorophore covalently attached to this domain revealed that, in the absence of ligands, this domain oscillates around a single detectable conformation. Binding to an antagonist does not change this conformation but does reduce the flexibility of the domain. However, when the β2 adrenergic receptor is bound to a full agonist, the G protein coupling domain exists in two distinct conformations. Moreover, the conformations induced by a full agonist can be distinguished from those induced by partial agonists. These results provide new insight into the structural consequence of antagonist binding and the basis of agonism and partial agonism.",
author = "Pejman Ghanouni and Zygmunt Gryczynski and Steenhuis, {Jacqueline J.} and Lee, {Tae Weon} and Farrens, {David L.} and Lakowicz, {Joseph R.} and Kobilka, {Brian K.}",
year = "2001",
month = "7",
day = "6",
doi = "10.1074/jbc.C100162200",
language = "English",
volume = "276",
pages = "24433--24436",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "27",

}

Functionally Different Agonists Induce Distinct Conformations in the G Protein Coupling Domain of the β2 Adrenergic Receptor. / Ghanouni, Pejman; Gryczynski, Zygmunt; Steenhuis, Jacqueline J.; Lee, Tae Weon; Farrens, David L.; Lakowicz, Joseph R.; Kobilka, Brian K.

In: Journal of Biological Chemistry, Vol. 276, No. 27, 06.07.2001, p. 24433-24436.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Functionally Different Agonists Induce Distinct Conformations in the G Protein Coupling Domain of the β2 Adrenergic Receptor

AU - Ghanouni, Pejman

AU - Gryczynski, Zygmunt

AU - Steenhuis, Jacqueline J.

AU - Lee, Tae Weon

AU - Farrens, David L.

AU - Lakowicz, Joseph R.

AU - Kobilka, Brian K.

PY - 2001/7/6

Y1 - 2001/7/6

N2 - G protein-coupled receptors represent the largest class of drug discovery targets. Drugs that activate G protein-coupled receptors are classified as either agonists or partial agonists. To study the mechanism whereby these different classes of activating ligands modulate receptor function, we directly monitored ligand-induced conformational changes in the G protein-coupling domain of the β2 adrenergic receptor. Fluorescence lifetime analysis of a reporter fluorophore covalently attached to this domain revealed that, in the absence of ligands, this domain oscillates around a single detectable conformation. Binding to an antagonist does not change this conformation but does reduce the flexibility of the domain. However, when the β2 adrenergic receptor is bound to a full agonist, the G protein coupling domain exists in two distinct conformations. Moreover, the conformations induced by a full agonist can be distinguished from those induced by partial agonists. These results provide new insight into the structural consequence of antagonist binding and the basis of agonism and partial agonism.

AB - G protein-coupled receptors represent the largest class of drug discovery targets. Drugs that activate G protein-coupled receptors are classified as either agonists or partial agonists. To study the mechanism whereby these different classes of activating ligands modulate receptor function, we directly monitored ligand-induced conformational changes in the G protein-coupling domain of the β2 adrenergic receptor. Fluorescence lifetime analysis of a reporter fluorophore covalently attached to this domain revealed that, in the absence of ligands, this domain oscillates around a single detectable conformation. Binding to an antagonist does not change this conformation but does reduce the flexibility of the domain. However, when the β2 adrenergic receptor is bound to a full agonist, the G protein coupling domain exists in two distinct conformations. Moreover, the conformations induced by a full agonist can be distinguished from those induced by partial agonists. These results provide new insight into the structural consequence of antagonist binding and the basis of agonism and partial agonism.

UR - http://www.scopus.com/inward/record.url?scp=0035816704&partnerID=8YFLogxK

U2 - 10.1074/jbc.C100162200

DO - 10.1074/jbc.C100162200

M3 - Article

C2 - 11320077

AN - SCOPUS:0035816704

VL - 276

SP - 24433

EP - 24436

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 27

ER -