Palmitoylation is required for signaling functions and membrane attachment of Gqα and Gsα

Philip B. Wedegaertner, David H. Chu, Paul T. Wilson, Mark J Levis, Henry R. Bourne

Research output: Contribution to journalArticle

Abstract

We have identified the palmitoylated cysteine residues of αq and αs, α subunits of two heterotrimeric G proteins. Mutational substitutions of serines for cysteines 9 and 10 in αq and cysteine 3 in αs profoundly alter behavior of the subunits expressed in HEK293 cells. Neither mutant α subunit incorporates palmitate; both mutant proteins are found in the soluble rather than the particulate fraction; mutant αq or αs cannot couple a co-expressed receptor to stimulation of phospholipase C or adenylylcyclase, respectively; cysteine substitution prevents a mutationally activated αq (R183C) from stimulating phospholipase C directly, and reduces but does not abolish the ability of a similarly activated αs (R201C) to stimulate cAMP synthesis. Substitution of a myristoylation sequence for the palmitoylation sites leads to labeling of αq and αs by myristate, rather than by palmitate. Myristoylation restores the abilities of both nonpalmitoylated αq and αs to attach to membranes and, in the case of αq, restores its ability to stimulate phospholipase C, whether triggered by the R183C mutation or by receptor activation. These findings identify palmitoylation as a critical determinant of membrane attachment for αq and αs and show that this modification is required for normal signaling by these proteins.

Original languageEnglish (US)
Pages (from-to)25001-25008
Number of pages8
JournalJournal of Biological Chemistry
Volume268
Issue number33
StatePublished - Nov 25 1993
Externally publishedYes

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Lipoylation
Cysteine
Type C Phospholipases
Membranes
Substitution reactions
Palmitates
Heterotrimeric GTP-Binding Proteins
HEK293 Cells
Myristic Acid
Mutant Proteins
Labeling
Serine
Chemical activation
Mutation
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Wedegaertner, P. B., Chu, D. H., Wilson, P. T., Levis, M. J., & Bourne, H. R. (1993). Palmitoylation is required for signaling functions and membrane attachment of Gqα and Gsα. Journal of Biological Chemistry, 268(33), 25001-25008.

Palmitoylation is required for signaling functions and membrane attachment of Gqα and Gsα. / Wedegaertner, Philip B.; Chu, David H.; Wilson, Paul T.; Levis, Mark J; Bourne, Henry R.

In: Journal of Biological Chemistry, Vol. 268, No. 33, 25.11.1993, p. 25001-25008.

Research output: Contribution to journalArticle

Wedegaertner, PB, Chu, DH, Wilson, PT, Levis, MJ & Bourne, HR 1993, 'Palmitoylation is required for signaling functions and membrane attachment of Gqα and Gsα', Journal of Biological Chemistry, vol. 268, no. 33, pp. 25001-25008.
Wedegaertner, Philip B. ; Chu, David H. ; Wilson, Paul T. ; Levis, Mark J ; Bourne, Henry R. / Palmitoylation is required for signaling functions and membrane attachment of Gqα and Gsα. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 33. pp. 25001-25008.
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