Covalently bound myristate in a lymphoma tyrosine protein kinase.
The murine lymphoma cell line LSTRA expresses high levels of a membrane-associated tyrosine protein kinase, which we now show to be acylated by [3H]myristate in vivo. This [3H]myristate-labeled tyrosine protein kinase is immunoprecipitated from detergent extracts of postnuclear particulate fractions with an antibody directed against its single site of tyrosine phosphorylation. This site has an amino acid sequence also found in the transforming proteins of the Rous sarcoma and Y73 viruses. Preincubation of the antibody with a peptide containing the same sequence completely blocks this immunoprecipitation. The [3H]myristate linkage to the protein is stable in boiling 2% NaDodSO4/0.125 M Tris Cl, pH 6.7/5% 2-mercaptoethanol, which suggests an amide rather than an ester linkage. Analogous attempts to label with [3H]palmitate show negligible incorporation into either nonnuclear particulate proteins or immunoprecipitated proteins. Chemical characterization of the immunoprecipitated protein isolated by NaDodSO4/PAGE verifies that the 3H label is in protein-associated myristate. Sonicated 5% NaDodSO4 extracts of LSTRA and YAC-1 (another murine lymphoma line) cells contain quite different distributions of myristoylated proteins.
D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008223:Lymphoma / Q000201:enzymology*; D051379:Mice; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism*; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases; D014316:Tritium
D009227:Myristic Acids; D010169:Palmitic Acids; D019814:Myristic Acid; D014316:Tritium; D019308:Palmitic Acid; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases