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

Journal Article · Mon Jan 01 04:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.2.728· OSTI ID:6709888

Mumby, S M; Gilman, A G ^[1]; Heukeroth, R O; Gordon, J I ^[2]

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1. Univ. of Texas Southwestern Medical Center, Dallas (USA)
2. Washington Univ. School of Medicine, St. Louis, MO (USA)

Myristolyation of seven different {alpha} 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 ({sup 3}H)myristate was incorporated into {alpha}{sub i1}, {alpha}{sub i2}, {alpha}{sub i3}, {alpha}{sub 0}, and {alpha}{sub 1}, and {alpha}{sub z} but not {alpha}{sub s} subunits. The role of myristoylation in the association of {alpha} 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 {alpha}{sub 0} 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 {alpha} 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 {alpha} subunits dissociated from {beta}{gamma}.

• OSTI ID: 6709888
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Journal Issue: 2 Vol. 87:2; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACYLATION
AMINES
AMINO ACIDS
AROMATICS
AZAARENES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
DISTRIBUTION
GLYCINE
GUANINE
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
MEMBRANE PROTEINS
MEMBRANES
MONOCARBOXYLIC ACIDS
MUTAGENESIS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PROTEINS
PURINES
SUBCELLULAR DISTRIBUTION
TETRADECANOIC ACID
TRITIUM COMPOUNDS
UPTAKE