Decay of an active GPCR

Conformational dynamics govern agonist rebinding and persistence of an active, yet empty, receptor state

Christopher T. Schafer, Jonathan F. Fay, Jay M. Janz, David Farrens

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Here, we describe two insights into the role of receptor conformational dynamics during agonist release (all-trans retinal, ATR) from the visual G protein-coupled receptor (GPCR) rhodopsin. First, we show that, after light activation, ATR can continually release and rebind to any receptor remaining in an active-like conformation. As with other GPCRs, we observe that this equilibrium can be shifted by either promoting the active-like population or increasing the agonist concentration. Second, we find that during decay of the signaling state an active-like, yet empty, receptor conformation can transiently persist after retinal release, before the receptor ultimately collapses into an inactive conformation. The latter conclusion is based on time-resolved, site-directed fluorescence labeling experiments that show a small, but reproducible, lag between the retinal leaving the protein and return of transmembrane helix 6 (TM6) to the inactive conformation, as determined from tryptophan-induced quenching studies. Accelerating Schiff base hydrolysis and subsequent ATR dissociation, either by addition of hydroxylamine or introduction of mutations, further increased the time lag between ATR release and TM6 movement. These observations show that rhodopsin can bind its agonist in equilibrium like a traditional GPCR, provide evidence that an active GPCR conformation can persist even after agonist release, and raise the possibility of targeting this key photoreceptor protein by traditional pharmaceutical-based treatments.

Original languageEnglish (US)
Pages (from-to)11961-11966
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number42
DOIs
StatePublished - Oct 18 2016

Fingerprint

G-Protein-Coupled Receptors
Rhodopsin
Hydroxylamine
Schiff Bases
Tryptophan
Proteins
Hydrolysis
Fluorescence
Light
Mutation
Pharmaceutical Preparations
Population

Keywords

  • Conformational dynamics
  • Fluorescence
  • GPCR
  • Retinal
  • Rhodopsin

ASJC Scopus subject areas

  • General

Cite this

Decay of an active GPCR : Conformational dynamics govern agonist rebinding and persistence of an active, yet empty, receptor state. / Schafer, Christopher T.; Fay, Jonathan F.; Janz, Jay M.; Farrens, David.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, No. 42, 18.10.2016, p. 11961-11966.

Research output: Contribution to journalArticle

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