Binding of regulator of G protein signaling (RGS) proteins to phospholipid bilayers. Contribution of location and/or orientation to GTPase-activating protein activity

Yaping Tu, Jimmy Woodson, Elliott M. Ross

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Regulator of G protein signaling (RGS) proteins must bind membranes in an orientation that permits the protein-protein interactions necessary for regulatory activity. RGS4 binds to phospholipid surfaces in a slow, multistep process that leads to maximal GTPase-activating protein (GAP) activity. When RGS4 is added to phospholipid vesicles that contain m2 or m1 muscarinic receptor and Gi, Gz, or Gq, GAP activity increases ∼3-fold over 4 h at 30 °C and more slowly at 20 °C. This increase in GAP activity is preceded by several other events that suggest that, after binding, optimal interaction with G protein and receptor requires reorientation of RGS4 on the membrane surface, a conformational change, or both. Binding of RGS4 is initially reversible but becomes irreversible within 5 min. Onset of irreversibility parallels initial quenching of tryptophan fluorescence (t1/2 ∼ 30 s). Further quenching occurs after binding has become irreversible (t1/2 ∼ 6 min) but is complete well before maximal GAP activity is attained. These processes all appear to be energetically driven by the amphipathic N-terminal domain of RGS4 and are accelerated by palmitoylation of cysteine residues in this region. The RGS4 N-terminal domain confers similar membrane binding behavior on the RGS domains of either RGS10 or RGSZ1.

Original languageEnglish
Pages (from-to)20160-20166
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number23
DOIs
StatePublished - Jun 8 2001

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RGS Proteins
GTPase-Activating Proteins
Phospholipids
Membranes
GTP-Binding Proteins
Quenching
GTP-Binding Protein Regulators
Muscarinic M2 Receptors
Muscarinic M1 Receptors
Lipoylation
Tryptophan
Cysteine
Proteins
Fluorescence

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Binding of regulator of G protein signaling (RGS) proteins to phospholipid bilayers. Contribution of location and/or orientation to GTPase-activating protein activity. / Tu, Yaping; Woodson, Jimmy; Ross, Elliott M.

In: Journal of Biological Chemistry, Vol. 276, No. 23, 08.06.2001, p. 20160-20166.

Research output: Contribution to journalArticle

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