The magnitude of the light-induced conformational change in different rhodopsins correlates with their ability to activate G proteins

Hisao Tsukamoto, David Farrens, Mitsumasa Koyanagi, Akihisa Terakita

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Light converts rhodopsin, the prototypical G protein-coupled receptor, into a form capable of activating G proteins. Recent work has shown that the light-activated state of different rhodopsins can possess different molecular properties, especially different abilities to activate G protein. For example, bovine rhodopsin is ∼20-fold more effective at activating G protein than parapinopsin, a non-visual rhodopsin, although these rhodopsins share relatively high sequence similarity. Here we have investigated possible structural aspects that might underlie this difference. Using a site-directed fluorescence labeling approach, we attached the fluorescent probe bimane to cysteine residues introduced in the cytoplasmic ends of transmembrane helices V and VI in both rhodopsins. The fluorescence spectra of these probes as well as their accessibility to aqueous quenching agents changed dramatically upon photoactivation in bovine rhodopsin but only moderately so in parapinopsin. We also compared the relative movement of helices V and VI upon photoactivation of both rhodopsins by introducing a bimane label and the bimane-quenching residue tryptophan into helices VI and V, respectively. Both receptors showed movement in this region upon activation, although the movement appears much greater in bovine rhodopsin than in parapinopsin. Together, these data suggest that a larger conformational change in helices V and VI of bovine rhodopsin explains why it has greater G protein activation ability than other rhodopsins. The different amplitude of the helix movement may also be responsible for functional diversity of G protein-coupled receptors.

Original languageEnglish (US)
Pages (from-to)20676-20683
Number of pages8
JournalJournal of Biological Chemistry
Volume284
Issue number31
DOIs
StatePublished - Jul 31 2009

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Rhodopsin
GTP-Binding Proteins
Light
G-Protein-Coupled Receptors
Quenching
Fluorescence
Chemical activation
Fluorescent Dyes
Tryptophan
Labeling
Cysteine
Labels

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

The magnitude of the light-induced conformational change in different rhodopsins correlates with their ability to activate G proteins. / Tsukamoto, Hisao; Farrens, David; Koyanagi, Mitsumasa; Terakita, Akihisa.

In: Journal of Biological Chemistry, Vol. 284, No. 31, 31.07.2009, p. 20676-20683.

Research output: Contribution to journalArticle

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