A truncated v-abl-derived tyrosine-specific tyrosine kinase expressed in Escherichia coli

M. L. Pritchard, D. Rieman, J. Feild, C. Kruse, M. Rosenberg, G. Poste, R. G. Greig, B. Q. Ferguson

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11 Scopus citations


Several biochemical properties of a 43 kDa v-abl-encoded tyrosine-specific protein kinase (p43(v-abl)) expressed in Escherichia coli were examined. p43(v-abl) is a fragment of a 60 kDa v-abl-encoded precursor, p60(v-abl), and could be generated by limited proteolysis of a purified p60(v-abl) with trypsin. Tryptic cleavage of p60(v-abl) was prevented in the presence of ATP. These results suggest that the catalytic kinase domain of v-abl-derived protein can be separated from other (regulatory) domains by limited proteolysis. p43(v-abl) readily phosphorylated tyrosine residues on several different protein and peptide substrates, including peptides containing only two amino acid residues. However, the local sequence of the tyrosine-containing peptide substrate significantly affected its rate of phosphorylation. Thus the primary structure and local conformation at the tyrosine acceptor site can play an important role in determining the substrate specificity of v-abl-derived kinase. Phosphorylation by p43(v-abl) requires Mn2+, Co2+ or Mg2+ and exhibits a strong preference for ATP as phosphate donor. Analogues of ATP and the thiol-reactive reagent N-ethylmaleimide inhibited p43(v-abl) kinase activity. Purified p43(v-abl) is intrinsically thermolabile (t( 1/2 ) = 5 min at 40°C) and phosphorylates glycerol inefficiently (K(m) = 1.4 M).

Original languageEnglish (US)
Pages (from-to)321-329
Number of pages9
JournalBiochemical Journal
Issue number2
StatePublished - 1989
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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