Palmitoylation of multiple Src-family kinases at a homologous N-terminal motif

M. Koegl, P. Zlatkine, S. C. Ley, Sara Courtneidge, A. I. Magee

Research output: Contribution to journalArticle

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Abstract

We have recently identified a novel N-terminal cysteine-containing motif which specifies the palmitoylation of several G-protein α-subunits. A related motif occurs at the N-terminus of members of the Src family of protein tyrosine kinases except for Src itself and Blk. We have investigated whether the Src, Fyn, Yes and Lck gene products are palmitoylated. Src was not labelled with [3H]palmitate when endogenously expressed in COS cells. In contrast, endogenous Yes immunoprecipitated from COS cells was palmitoylated. Fyn was palmitoylated in insect cells infected with a recombinant baculovirus and the palmitoylation was independent of protein synthesis, suggesting a dynamic turnover of this lipid. Fatty acid analysis indicated that most of the label was incorporated as palmitate. Lck was palmitoylated when expressed by transfection in COS cells. All of these protein tyrosine kinases were also detectably myristoylated in each of the systems tested. Experiments performed with mutants of Lck expressed by transfection in COS cells indicated that cysteines at positions 3 and 5 were both palmitoylation sites and that myristoylation was required for palmitoylation. To confirm that palmitoylation was occurring on cysteines in the N-terminal region of Fyn, site-directed mutagenesis was used to replace the cysteines at positions 3 and 6 with alanine. The resulting protein was not palmitoylated but was still myristoylated when expressed in COS cells. A glycine to alanine mutant at position 2 was also not palmitoylated, showing that myristoylation is a prerequisite for palmitoylation. Our data indicate that Src family members containing the N-terminal cysteine motif are indeed palmitoylated. By analogy with Ras, it is possible that palmitoylation may play an important role in the localization and function of Src family protein tyrosine kinases.

Original languageEnglish (US)
Pages (from-to)749-753
Number of pages5
JournalBiochemical Journal
Volume303
Issue number3
StatePublished - 1994
Externally publishedYes

Fingerprint

Lipoylation
src-Family Kinases
Cysteine
COS Cells
Palmitates
Alanine
Protein-Tyrosine Kinases
Transfection
Mutagenesis
Protein Subunits
GTP-Binding Proteins
Glycine
Labels
Baculoviridae
Proteins
Fatty Acids
Genes
Site-Directed Mutagenesis
Lipids
Insects

ASJC Scopus subject areas

  • Biochemistry

Cite this

Koegl, M., Zlatkine, P., Ley, S. C., Courtneidge, S., & Magee, A. I. (1994). Palmitoylation of multiple Src-family kinases at a homologous N-terminal motif. Biochemical Journal, 303(3), 749-753.

Palmitoylation of multiple Src-family kinases at a homologous N-terminal motif. / Koegl, M.; Zlatkine, P.; Ley, S. C.; Courtneidge, Sara; Magee, A. I.

In: Biochemical Journal, Vol. 303, No. 3, 1994, p. 749-753.

Research output: Contribution to journalArticle

Koegl, M, Zlatkine, P, Ley, SC, Courtneidge, S & Magee, AI 1994, 'Palmitoylation of multiple Src-family kinases at a homologous N-terminal motif', Biochemical Journal, vol. 303, no. 3, pp. 749-753.
Koegl, M. ; Zlatkine, P. ; Ley, S. C. ; Courtneidge, Sara ; Magee, A. I. / Palmitoylation of multiple Src-family kinases at a homologous N-terminal motif. In: Biochemical Journal. 1994 ; Vol. 303, No. 3. pp. 749-753.
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