Functional analysis of protein N-myristoylation: Metabolic labeling studies using three oxygen-substituted analogs of myristic acid and cultured mammalian cells provide evidence for protein-sequence-specific incorporation and analog-specific redistribution

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

DOI:https://doi.org/10.1073/pnas.87.21.8511· OSTI ID:5603336

Johnson, D R; Heuckeroth, R O; Gordon, J I ^[1]; Cox, A D; Solski, P A; Buss, J E ^[2]; Devadas, B; Adams, S P; Leimgruber, R M ^[3]

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1. Washington Univ., St. Louis, MO (United States)
2. La Jolla Cancer Research Foundation, CA (United States)
3. Monsanto Co., St. Louis, MO (United States)

Covalent attachment of myristic acid (C14:0) to the NH{sub 2}-terminal glycine residue of a number of cellular, viral, and oncogene-encoded proteins is essential for full expression of their biological function. Substitution of oxygen for methylene groups in this fatty acid does not produce a significant change in chain length or stereochemistry but does result in a reduction in hydrophobicity. These heteroatom-containing analogs serve as alternative substrates for mammalian myristoyl-CoA: protein N-myristoyltransferase and offer the opportunity to explore structure/function relationships of myristate in N-myristoyltransferase proteins. The authors have synthesized three tritiated analogs of myristate with oxygen substituted for methylene groups at C6, C11, and C13. Metabolic labeling studies were performed with these compounds and (i) a murine myocyte cell line (BC{sub 3}H1), (ii) a rat fibroblast cell that produces p60{sup v-src} (3Xsrc), or (iii) NIH 3T3 cells that have been engineered to express a fusion protein consisting of an 11-residue myristoylation signal from the Rasheed sarcoma virus (RaSV) gag protein linked to c-Ha-ras with a Cys {yields} Ser-186 mutation. Two-dimensional gel electrophoresis of membrane and soluble fractions prepared from cell lysates revealed different patterns of incorporation of the analogs into cellular N-myristoyl proteins. The demonstration that these analogs differ in the extent to which they are incorporated and in their ability to cause redistribution of any single protein suggests that they may also have sufficient selectivity to be of potential therapeutic value.

• OSTI ID: 5603336
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 87:21; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
BIOCHEMISTRY
CARBOXYLIC ACIDS
CHEMISTRY
DISTRIBUTION
ELECTROPHORESIS
GLYCINE
HYDROGEN COMPOUNDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
SUBCELLULAR DISTRIBUTION
TRITIUM COMPOUNDS