G-protein alpha-subunit expression, myristoylation, and membrane association in COS cells.
- 1 January 1990
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 87 (2) , 728-732
- https://doi.org/10.1073/pnas.87.2.728
Abstract
Myristoylation 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 [3H]myristate was incorporated into .alpha.i1, .alpha.i2, .alpha.i3, .alpha.0, .alpha.t, and .alpha.z but not .alpha.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.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 dissociate from .beta..gamma.This publication has 30 references indexed in Scilit:
- Specific and saturable binding of pp60v-src to plasma membranes: Evidence for a myristyl-src receptorCell, 1989
- Sticky fingers and CAAX boxesNature, 1988
- THE BIOLOGY AND ENZYMOLOGY OF EUKARYOTIC PROTEIN ACYLATIONAnnual Review of Biochemistry, 1988
- GTP analogues cause release of the alpha subunit of the GTP binding protein, GO, from the plasma membrane of NG108-15 cellsBiochemical and Biophysical Research Communications, 1988
- Myristoylated alpha subunits of guanine nucleotide-binding regulatory proteins.Proceedings of the National Academy of Sciences, 1987
- Hydroxylamine-stable covalent linkage of myristic acid in Goα, a guanine nucleotide-binding protein of bovine brainBiochemical and Biophysical Research Communications, 1987
- G PROTEINS: TRANSDUCERS OF RECEPTOR-GENERATED SIGNALSAnnual Review of Biochemistry, 1987
- Purification and characterization of yeast myristoyl CoA:protein N-myristoyltransferase.Proceedings of the National Academy of Sciences, 1987
- Oligonucleotide-Directed Mutagenesis: A Simple Method Using Two Oligonucleotide Primers and a Single-Stranded DNA TemplateDNA, 1984
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976