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PMID: 6595656 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Covalently bound myristate in a lymphoma tyrosine protein kinase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 24 ·1984-12-00 ·Pages 7679-82

Marchildon GA, Casnellie JE, Walsh KA, Krebs EG

Abstract

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.

MeSH Terms
Animals Cell Line Chromatography, High Pressure Liquid Electrophoresis, Polyacrylamide Gel Lymphoma/enzymology Mice Myristic Acid Myristic Acids/metabolism Palmitic Acid Palmitic Acids/metabolism Protein Binding Protein Kinases/metabolism Protein-Tyrosine Kinases Tritium
Chemicals
Myristic Acids Palmitic Acids Myristic Acid Tritium Palmitic Acid Protein Kinases Protein-Tyrosine Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Marchildon G A
Casnellie J E
Walsh K A
Krebs E G
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.24.7679
Published
1984-12-00
Pages
7679-82
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC392215
Subset
IM
Grants
NIGMS NIH HHS · GM 07270 · United States
NIGMS NIH HHS · GM 15731 · United States
External Links
PubMed source DOI: 10.1073/pnas.81.24.7679 PMC Full Text View journal details
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