Receptor-mediated activation of heterotrimeric G-proteins in living cells

C. Janetopoulos, T. Jin, P. Devreotes

Research output: Contribution to journalArticlepeer-review

Abstract

Receptor-mediated activation of heterotrimeric GTP-binding proteins (G-proteins) was visualized in living Dictyostelium discoideum cells by monitoring fluorescence resonance energy transfer (FRET) between α- and β- subunits fused to cyan and yellow fluorescent proteins. The G-protein heterotrimer rapidly dissociated and reassociated upon addition and removal of chemoattractant. During continuous stimulation, G-protein activation reached a dose-dependent steady-state level. Even though physiological responses subsided, the activation did not decline. Thus, adaptation occurs at another point in the signaling pathway, and occupied receptors, whether or not they are phosphorylated, catalyze the G-protein cycle. Construction of similar energy-transfer pairs of mammalian G-proteins should enable direct in situ mechanistic studies and applications such as drug screening and identifying ligands of newly found G-protein-coupled receptors.

Original languageEnglish (US)
Pages (from-to)2408-2411
Number of pages4
JournalScience
Volume291
Issue number5512
DOIs
StatePublished - Mar 23 2001

ASJC Scopus subject areas

  • General

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