title: Detection of c-abl tyrosine kinase activity in vitro permits direct comparison of normal and altered abl gene products.
abstract: The v-abl transforming protein P160v-abl and the P210c-abl gene product of the translocated c-abl gene in Philadelphia chromosome-positive chronic myelogenous leukemia cells have tyrosine-specific protein kinase activity. Under similar assay conditions the normal c-abl gene products, murine P150c-abl and human P145c-abl, lacked detectable kinase activity. Reaction conditions were modified to identify conditions which would permit the detection of c-abl tyrosine kinase activity. It was found that the Formalin-fixed Staphylococcus aureus formerly used for immunoprecipitation inhibits in vitro abl kinase activity. In addition, the sodium dodecyl sulfate and deoxycholate detergents formerly used in the cell lysis buffer were found to decrease recovered abl kinase activity. The discovery of assay conditions for c-abl kinase activity now makes it possible to compare P150c-abl and P145c-abl kinase activity with the altered abl proteins P160v-abl and P210c-abl. Although all of the abl proteins have in vitro tyrosine kinase activity, they differ in the way they utilize themselves as substrates in vitro. Comparison of in vitro and in vivo tyrosine phosphorylation sites of the abl proteins suggests that they function differently in vivo. The development of c-abl kinase assay conditions should be useful in elucidating c-abl function.
mesh_terms: D000011:Abelson murine leukemia virus / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D005796:Genes; D006801:Humans; D007951:Leukemia, Myeloid / Q000235:genetics; D051379:Mice; D009154:Mutation; D010677:Philadelphia Chromosome; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D013945:Thymoma; D013953:Thymus Neoplasms; D014178:Translocation, Genetic; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D014764:Viral Proteins; D011505:Protein-Tyrosine Kinases