Distinct structural changes in a G protein-coupled receptor caused by different classes of agonist ligands.
The activity of G protein-coupled receptors can be modulated by different classes of ligands, including agonists that promote receptor signaling and inverse agonists that reduce basal receptor activity. The conformational changes in receptor structure induced by different agonist ligands are not well understood at present. In this study, we employed an in situ disulfide cross-linking strategy to monitor ligand-induced conformational changes in a series of cysteine-substituted mutant M(3) muscarinic acetylcholine receptors. The observed disulfide cross-linking patterns indicated that muscarinic agonists trigger a separation of the N-terminal segment of the cytoplasmic tail (helix 8) from the cytoplasmic end of transmembrane domain I. In contrast, inverse muscarinic agonists were found to increase the proximity between these two receptor regions. These findings provide a structural basis for the opposing biological effects of muscarinic agonists and inverse agonists. This study also provides the first piece of direct structural information as to how the conformations induced by these two functionally different classes of ligands differ at the molecular level. Given the high degree of structural homology found among most G protein-coupled receptors, our findings should be of broad general relevance.
Li
Jian Hua
JH
Molecular Signaling Section, Laboratory of Bioorganic Chemistry, NIDDK, National Institutes of Health, Bethesda, Maryland 20892-0810, USA.
Han
Sung-Jun
SJ
Hamdan
Fadi F
FF
Kim
Soo-Kyung
SK
Jacobson
Kenneth A
KA
Bloodworth
Lanh M
LM
Zhang
Xiaohong
X
Wess
Jürgen
J
D019943:Amino Acid Substitution; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003545:Cysteine / Q000235:genetics; D008024:Ligands; D008958:Models, Molecular; D020125:Mutation, Missense; D017433:Protein Structure, Secondary / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D051381:Rats; D043587:Receptor, Muscarinic M3 / Q000819:agonists*
D008024:Ligands; D043587:Receptor, Muscarinic M3; D003545:Cysteine