G-protein α-subunit expression, myristoylation, and membrane association in COS cells

Susanne M. Mumby, Robert O. Heukeroth, Jeffrey I. Gordon, Alfred G. Gilman

Research output: Contribution to journalArticle

205 Citations (Scopus)

Abstract

Myristoylation of seven different a subunits of guanine nucleotide-binding regulatory proteins (G proteins) was examined by expressing these proteins in monkey kidney COS cells. Metabolic labeling studies of cells transfected with cytomegalovirus-based expression vectors indicated that [3H]myristate was incorporated into αi2, αi2, αi3, α0, αt, and αz, but not αs subunits. The role of myristoylation in the association of α subunits with membranes was analyzed by site-directed mutagenesis and by substitution of myristate with a less hydrophobic analog, 10-(propoxy)decanoate (11-oxamyristate). Myristoylation of α0 was blocked when an alanine residue was substituted for its amino-terminal glycine, as was association of the protein with membranes. Substitution of the myristoyl group with 11-oxamyristate affected the cellular distribution of a subset of acylated a subunits. The results are consistent with a model wherein the hydrophobic interaction of myristate with the bilayer permits continued association of the protein with the plasma membrane when G-protein α subunits dissociate from βγ.

Original languageEnglish (US)
Pages (from-to)728-732
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number2
StatePublished - 1990

Fingerprint

COS Cells
Protein Subunits
Myristic Acid
GTP-Binding Proteins
Membranes
Decanoates
Site-Directed Mutagenesis
Cytomegalovirus
Hydrophobic and Hydrophilic Interactions
Alanine
Glycine
Haplorhini
Carrier Proteins
Membrane Proteins
Proteins
Cell Membrane
Kidney

Keywords

  • Fatty acid analog
  • Protein N-myristoylation

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

G-protein α-subunit expression, myristoylation, and membrane association in COS cells. / Mumby, Susanne M.; Heukeroth, Robert O.; Gordon, Jeffrey I.; Gilman, Alfred G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 2, 1990, p. 728-732.

Research output: Contribution to journalArticle

Mumby, Susanne M. ; Heukeroth, Robert O. ; Gordon, Jeffrey I. ; Gilman, Alfred G. / G-protein α-subunit expression, myristoylation, and membrane association in COS cells. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 2. pp. 728-732.
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