Abstract
Protein prenylation with farnesyl or geranylgeranyl moieties is an important posttranslational modification that affects the activity of such diverse proteins as the nuclear lamins, the yeast mating factor mata, and the ras oncogene products. In this article, we show that whole retinal cultures incorporate radioactive mevalonic acid into proteins of 23-26 kDa and one of 8 kDa. The former proteins are probably the "small" guanine nucleotide-binding regulatory proteins (G proteins) and the 8-kDa protein is the gamma subunit of the well-studied retinal heterotrimeric G protein (transducin). After deprenylating purified transducin and its subunits with Raney nickel or methyl iodide/base, the adducted prenyl group can be identified as an all-trans-farnesyl moiety covalently linked to a cysteine residue. Thus far, prenylation reactions have been found to occur at cysteine in a carboxyl-terminal consensus CAAX sequence, where C is the cysteine, A is an aliphatic amino acid, and X is undefined. Both the alpha and gamma subunits of transducin have this consensus sequence, but only the gamma subunit is prenylated. Therefore, the CAAX motif is not necessary and sufficient to direct prenylation. Finally, since transducin is the best understood G protein, both structurally and mechanistically, the discovery that it is farnesylated should allow for a quantitative understanding of this post-translational modification.
MeSH Terms
Animals
Cattle
Diterpenes/analysis
Farnesol/analysis
Gas Chromatography-Mass Spectrometry
In Vitro Techniques
Leucine/metabolism
Macromolecular Substances
Mevalonic Acid/metabolism
Protein Processing, Post-Translational
Retina/metabolism
Transducin/biosynthesis,chemistry,genetics
Chemicals
Diterpenes
Macromolecular Substances
Farnesol
geranylgeraniol
Transducin
Leucine
Mevalonic Acid
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lai R K
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Perez-Sala D
Cañada F J
Rando R R
References (31)
31 references, click to expand
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
Farnesylated gamma-subunit of photoreceptor G protein indispensable for GTP-binding.
Nature. 1990 Aug 16;346(6285):658-60
PMID: 2385292
-
Feedback regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in livers of mice treated with mevinolin, a competitive inhibitor of the reductase.
J Clin Invest. 1980 Nov;66(5):1094-100
PMID: 6903572
-
A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids.
Anal Biochem. 1984 Apr;138(1):141-3
PMID: 6731838
-
Isolation and characterization of a cDNA clone for the gamma subunit of bovine retinal transducin.
Proc Natl Acad Sci U S A. 1984 Nov;81(22):6948-52
PMID: 6438626
-
Primary structure of the alpha-subunit of transducin and its relationship to ras proteins.
Nature. 1985 May 16-22;315(6016):242-5
PMID: 3923359
-
Amino acid sequence of the alpha subunit of transducin deduced from the cDNA sequence.
Proc Natl Acad Sci U S A. 1985 Jul;82(13):4311-5
PMID: 2409555
-
GTPase of bovine rod outer segments: the amino acid sequence of the alpha subunit as derived from the cDNA sequence.
Proc Natl Acad Sci U S A. 1985 Jul;82(13):4316-20
PMID: 2989813
-
Pertussis toxin-catalyzed ADP-ribosylation of transducin. Cysteine 347 is the ADP-ribose acceptor site.
J Biol Chem. 1985 Nov 25;260(27):14428-30
PMID: 3863818
-
Reversible staining and peptide mapping of proteins transferred to nitrocellulose after separation by sodium dodecylsulfate-polyacrylamide gel electrophoresis.
Anal Biochem. 1986 Aug 1;156(2):341-7
PMID: 2429581
-
Transducin interactions with rhodopsin. Evidence for positive cooperative behavior.
J Biol Chem. 1987 Sep 15;262(26):12444-7
PMID: 3114258
-
Isoprenylated proteins in cultured cells: subcellular distribution and changes related to altered morphology and growth arrest induced by mevalonate deprivation.
J Cell Physiol. 1987 Dec;133(3):471-81
PMID: 3693410
-
Competitive inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase by ML-236A and ML-236B fungal metabolites, having hypocholesterolemic activity.
FEBS Lett. 1976 Dec 31;72(2):323-6
PMID: 16386050
-
Incorporation of a product of mevalonic acid metabolism into proteins of Chinese hamster ovary cell nuclei.
J Cell Biol. 1988 Oct;107(4):1307-16
PMID: 3170631
-
Structure of Saccharomyces cerevisiae mating hormone a-factor. Identification of S-farnesyl cysteine as a structural component.
J Biol Chem. 1988 Dec 5;263(34):18236-40
PMID: 3056940
-
Molecular mechanism of visual transduction.
Eur J Biochem. 1989 Feb 1;179(2):255-66
PMID: 2537204
-
Beta gamma-subunit of bovine transducin composed of two components with distinctive gamma-subunits.
J Biol Chem. 1989 Apr 5;264(10):5937-43
PMID: 2925642
-
All ras proteins are polyisoprenylated but only some are palmitoylated.
Cell. 1989 Jun 30;57(7):1167-77
PMID: 2661017
-
Enzymatic methylation of 23-29-kDa bovine retinal rod outer segment membrane proteins. Evidence for methyl ester formation at carboxyl-terminal cysteinyl residues.
J Biol Chem. 1989 Aug 5;264(22):12879-84
PMID: 2753892
-
Molecular basis of visual excitation.
Cold Spring Harb Symp Quant Biol. 1988;53 Pt 1:283-94
PMID: 3076083
-
Characterization of isoprenoid involved in the post-translational modification of mammalian cell proteins.
J Biol Chem. 1989 Oct 25;264(30):18168-72
PMID: 2808372
-
The conserved carboxy-terminal cysteine of nuclear lamins is essential for lamin association with the nuclear envelope.
J Cell Biol. 1989 Nov;109(5):2003-11
PMID: 2808518
-
p21ras is modified by a farnesyl isoprenoid.
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8323-7
PMID: 2682646
-
Human lamin B contains a farnesylated cysteine residue.
J Biol Chem. 1989 Dec 5;264(34):20422-9
PMID: 2684976
-
Prenylated proteins: the structure of the isoprenoid group.
Science. 1990 Jan 19;247(4940):318-20
PMID: 2296720
-
Identification of geranylgeranyl-modified proteins in HeLa cells.
Science. 1990 Jan 19;247(4940):320-2
PMID: 2296721
-
Post-translational modification of low molecular mass GTP-binding proteins by isoprenoid.
J Biol Chem. 1990 Feb 5;265(4):2148-55
PMID: 2105317
-
Regulation of the mevalonate pathway.
Nature. 1990 Feb 1;343(6257):425-30
PMID: 1967820
-
Prenyl proteins in eukaryotic cells: a new type of membrane anchor.
Trends Biochem Sci. 1990 Apr;15(4):139-42
PMID: 2187294
-
Fatty acylated proteins as components of intracellular signaling pathways.
Biochemistry. 1990 Mar 20;29(11):2623-34
PMID: 2189494
-
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
Anal Biochem. 1976 May 7;72:248-54
PMID: 942051