Assay of interferon-mediated protein kinase activity from plasma and tissue extracts.
Interferon-treated mouse and human cells show enhanced levels of a protein kinase activity which is manifested by the phosphorylation of endogenous 67,000 and 72,000 Mr proteins, respectively. Enhanced levels of such kinase activity are also detectable in the plasma of patients treated with interferon and in the plasma and tissues of interferon-treated mice. A rapid and efficient method of assay for these protein kinase activities is described. The samples are first incubated with heparin (100 units/ml), which results in the inhibition of different protein kinase activities, but not the one mediated by interferon. The latter one is then assayed after partial purification on poly(rI):(rC)-Sepharose or poly(rG)-Sepharose. The protein kinase from human and mouse cells in culture and from the different tissues of mice binds specifically to poly(rI):(rC)-Sepharose. On the other hand, the protein kinase activity from both mouse and human plasma shows a higher affinity toward poly(rG)-Sepharose. These methods are successfully applied for the determination of the interferon-mediated protein kinase activity from tissue extracts and plasma.
D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D006367:HeLa Cells; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D007739:L Cells; D051379:Mice; D011119:Polynucleotides / Q000378:metabolism; D011485:Protein Binding; D011494:Protein Kinases / Q000032:analysis* / Q000097:blood; D014020:Tissue Extracts / Q000032:analysis
D011119:Polynucleotides; D014020:Tissue Extracts; D006493:Heparin; D007372:Interferons; D011494:Protein Kinases