Characterization of casein kinase II from a virally transformed macrophage-like cell line, RAW264.
Casein kinase II from a virally-transformed macrophage cell line (RAW264) was purified by a sequential DEAE, Procion Red, phosvitin-Sepharose and heparin-Sepharose chromatography. With [tau-32P]GTP as a phosphate donor and casein as a substrate, the kinase was stimulated by polyamines and inhibited by heparin. The purified kinase had a specific activity of 1137 nmol/min/mg protein and exhibited three major protein bands of 40 K, 35 K, and 25 K. Under non-denaturing conditions in 50 mM Tris-50 mM NaCl the enzyme was eluted as a single peak with molecular weight of 110 K. Incubation of kinase in the presence of [tau-32P]GTP and Mg2+ resulted in phosphorylation of the 25 K protein band of the enzyme. In the presence of [tau-32P]GTP and Mg2+ the kinase was able to phosphorylate 55 K protein band in purified ornithine decarboxylase preparation from RAW264 cells and the rat-type II regulatory subunit of the cyclic AMP-dependent protein kinase.
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D000818:Animals; D047388:Casein Kinases; D002364:Caseins / Q000378:metabolism; D002461:Cell Line, Transformed; D006493:Heparin / Q000494:pharmacology; D008264:Macrophages / Q000201:enzymology*; D009955:Ornithine Decarboxylase / Q000378:metabolism; D010766:Phosphorylation; D011073:Polyamines / Q000494:pharmacology; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism; D011499:Protein Processing, Post-Translational
D002364:Caseins; D011073:Polyamines; D047428:Protein Kinase Inhibitors; D006493:Heparin; D011494:Protein Kinases; D047388:Casein Kinases; D009955:Ornithine Decarboxylase