Identification of the human T-lymphocyte protein-tyrosine kinase by peptide-specific antibodies.

We have recently cloned a cDNA from the human T-cell leukemia, JURKAT, having homology with the src-like family of protein-tyrosine kinases. We have made rabbit polyclonal antibodies against the synthetic peptide CKERPEDRPTFDYLRSVLEDFFTATEGQYQPQP (cys-33-pro) deduced from the carboxy-terminal amino acid sequence predicted by the JURKAT cDNA. In this report, we demonstrate that these antibodies immunoprecipitate the protein-tyrosine kinase activity from solubilized membrane extracts from JURKAT T-leukemia cells and from human peripheral blood T-lymphocytes from normal donors. A 58 kd protein, exhibiting protein-tyrosine kinase activity, was specifically immunoprecipitated in both cases. The antibodies failed to crossreact with pp60c-src from human platelets, but did crossreact with the murine T-lymphocyte protein-tyrosine kinase, pp56T-cell.