Biochemical characterization of a 34-kilodalton normal cellular substrate of pp60v-src and an associated 6-kilodalton protein.
Transformation of fibroblasts by several retroviruses that produce transforming gene products associated with protein kinase activity results in the phosphorylation of a normal cellular protein with an Mr of 34,000 (the 34K protein). Evidence is presented here that, as extracted from chicken embryo fibroblasts, this protein exists in two forms that differ both in their elution from hydroxylapatite and in their native molecular weight. The form that eluted from hydroxylapatite at 210 to 295 mM potassium phosphate displayed a native molecular weight of 30,000 to 40,000, whereas the form that eluted at 320 to 440 mM displayed a native molecular weight of 60,000 to 70,000. The latter form copurified with a low-molecular-weight protein with an approximate Mr of 6,000 (6K). Both forms of 34K were completely separable from malate dehydrogenase activity. Phosphorylated 34K, isolated from Rous sarcoma virus-transformed cells, was also present in two forms; hence, in the cell neither form serves as a preferential substrate for pp60v-src. We found that the expression of 34K differed greatly in various avian tissues. In particular, it was present in the highest concentration in cultured fibroblasts and in very low concentration in brain tissue. Its expression in this tissue seems to be controlled at the level of transcription, since 34K mRNA in brain tissue was barely detectable. The expression of 6K was similar to that of 34K.
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002472:Cell Transformation, Viral; D002642:Chick Embryo; D005347:Fibroblasts / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000302:isolation & purification; D008970:Molecular Weight; D015688:Oncogene Protein pp60(v-src); D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins / Q000302:isolation & purification*; D013379:Substrate Specificity; D014764:Viral Proteins / Q000378:metabolism*
D011506:Proteins; D014764:Viral Proteins; D008291:Malate Dehydrogenase; D011494:Protein Kinases; D015688:Oncogene Protein pp60(v-src)