Isolation of a catalytically competent phosphorylated tyrosine kinase from Rous sarcoma virus-induced rat tumor by immunoadsorption to and hapten elution from phosphotyrosine binding antibodies.
A procedure has been developed for the isolation of a catalytically competent phosphorylated tyrosine kinase (RSV Y-kinase) from avian sarcoma virus-induced rat tumors. The procedure involves reaction of partially purified RSV Y-kinase with ATP to effect tyrosyl phosphorylation of catalytically competent RSV Y-kinase. Tyrosyl phosphorylated RSV Y-kinase was isolated from the heterogeneous reaction mixture by immunoadsorption on immobilized phosphotyrosyl binding antibodies and elution with the hapten p-nitrophenyl phosphate. Estimation of the phosphate content of the purified phosphorylated RSV Y-kinase indicated that 1-3 tyrosyl groups had been phosphorylated upon reaction with ATP. The specific activity toward histone 2B of the purified phosphorylated RSV Y-kinase was at least 30-fold greater than that estimated for the RSV Y-kinase prepared previously by immunoadsorption on immobilized antiserum from tumor bearing rabbits.
Brain Tumor Research Center, University of California, San Francisco.
S
S
Whitehouse-Hills
L S
LS
Argetsinger
J A
JA
Shafer
D000818:Animals; D000906:Antibodies; D001358:Avian Sarcoma Viruses / Q000201:enzymology*; D006241:Haptens; D006657:Histones / Q000378:metabolism; D007163:Immunosorbent Techniques; D009374:Neoplasms, Experimental / Q000201:enzymology* / Q000209:etiology; D015688:Oncogene Protein pp60(v-src) / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases / Q000302:isolation & purification*; D051381:Rats; D011916:Rats, Inbred F344; D001357:Sarcoma, Avian; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000276:immunology
D000906:Antibodies; D006241:Haptens; D006657:Histones; D019000:Phosphotyrosine; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)