Partial purification and characterization of a pp60v-src-related tyrosine kinase from bovine brain.
We have identified and substantially purified a tyrosine protein kinase from normal bovine brain that is immunologically related to the product of the Rous sarcoma virus oncogene (pp60v-src). The enzyme, a 61-kDa protein (p61), is solubilized with detergent from bovine cerebral cortical membranes and purified by column chromatography. In the purest preparations, this protein is phosphorylated only on tyrosine, but it can also be a substrate for serine- and threonine-specific protein kinases. The p61 protein phosphorylates the heavy chain of immunoglobulins from rabbits bearing Rous sarcoma virus-induced tumors (TBR IgG) but not normal IgG. TBR IgG precipitates the 61-kDa phosphoprotein and protein kinase activity from purified preparations. The activity of the purified brain tyrosine kinase is 10 times higher in the presence of 7-10 mM Mn2+ and 6 mM Mg2+ than it is with 6 mM Mg2+ alone. With Mn2+, the p61 enzyme has a Km for ATP of 2 microM. All preparations of p61 also contain a 64-kDa protein (p64) that is phosphorylated on tyrosine. Measurement of the Stokes radius of p61 and p64 by gel filtration shows that they are not physically associated in buffer containing the nonionic detergent Lubrol 12A9. The p64 protein is not precipitated by TBR IgG. We do not know whether p64 is only a substrate for the p61 tyrosine kinase or is itself a kinase.
E J
EJ
Neer
J M
JM
Lok
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001358:Avian Sarcoma Viruses; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D002540:Cerebral Cortex / Q000201:enzymology; D007074:Immunoglobulin G / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008970:Molecular Weight; D015688:Oncogene Protein pp60(v-src); D010750:Phosphoproteins / Q000302:isolation & purification*; D019000:Phosphotyrosine; D011487:Protein Conformation; D011494:Protein Kinases / Q000302:isolation & purification*; D011505:Protein-Tyrosine Kinases; D013379:Substrate Specificity; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis; D014764:Viral Proteins / Q000302:isolation & purification
D007074:Immunoglobulin G; D010750:Phosphoproteins; D014764:Viral Proteins; D019000:Phosphotyrosine; D014443:Tyrosine; D008345:Manganese; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)