Inhibition of tyrosine protein kinases by halomethyl ketones

Javier Navarro, M. Abdel Ghany, E. Racker

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

A chloromethyl ketone derivative of lactic acid was shown to inhibit protein phosphorylation in plasma membranes of Ehrlich ascites tumor cells [Johnson, H. J., Zimniak, A., & Racker, E. (1982) Biochemistry 21, 2984-2989]. We now show that this inhibitor as well as three halomethyl ketone derivatives of amino acids and peptides specifically inhibits tyrosine protein kinase activity in intact plasma membranes and Triton extracts of plasma membrane of A-431 tumor cells. The most effective inhibitor is a bromomethyl ketone derivative of leucine that inhibits the phosphorylation of a protein that migrates to the same position as the EGF receptor in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Inhibition of phosphorylation took place in the presence or absence of added EGF, and the inhibitor did not interfere with the binding of EGF to the receptor nor with the dephosphorylation of the EGF-stimulated phosphoprotein. EGF-dependent phosphorylation in a Triton extract of plasma membranes from normal placenta was considerably less sensitive to the bromomethyl ketone derivative of leucine. The tyrosine protein kinase activity of the transformation gene product of Fujinami virus was particularly sensitive to the bromomethyl ketone derivative of leucine, while the src gene product of Rous sarcoma virus was comparatively less sensitive. The bromomethyl ketone inhibitor interfered with the phosphorylation of the EGF receptor by [γ-32P]-8-azido-ATP but much less with the light-sensitive binding. This observation and the lack of interference with EGF binding suggest that the inhibitor interacts with the protein kinase portion of the receptor complex.

Original languageEnglish (US)
Pages (from-to)6138-6144
Number of pages7
JournalBiochemistry
Volume21
Issue number24
StatePublished - 1982
Externally publishedYes

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Phosphorylation
Ketones
Protein-Tyrosine Kinases
Epidermal Growth Factor
Cell membranes
Epidermal Growth Factor Receptor
Cell Membrane
Leucine
Derivatives
Viruses
src Genes
Tumors
Rous sarcoma virus
Ehrlich Tumor Carcinoma
Genes
Cells
Phosphoproteins
Biochemistry
Sodium Dodecyl Sulfate
Protein Kinases

ASJC Scopus subject areas

  • Biochemistry

Cite this

Navarro, J., Ghany, M. A., & Racker, E. (1982). Inhibition of tyrosine protein kinases by halomethyl ketones. Biochemistry, 21(24), 6138-6144.

Inhibition of tyrosine protein kinases by halomethyl ketones. / Navarro, Javier; Ghany, M. Abdel; Racker, E.

In: Biochemistry, Vol. 21, No. 24, 1982, p. 6138-6144.

Research output: Contribution to journalArticle

Navarro, J, Ghany, MA & Racker, E 1982, 'Inhibition of tyrosine protein kinases by halomethyl ketones', Biochemistry, vol. 21, no. 24, pp. 6138-6144.
Navarro, Javier ; Ghany, M. Abdel ; Racker, E. / Inhibition of tyrosine protein kinases by halomethyl ketones. In: Biochemistry. 1982 ; Vol. 21, No. 24. pp. 6138-6144.
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