Cyclic nucleotide independent casein kinase (G type) in bovine adrenal cortex: purification and properties of two molecular forms.
Two soluble cyclic nucleotide independent protein kinase (ATP: protein-phosphotransferase, EC 2.7.1.37) activities have been purified from bovine adrenal cortex cytosol. Both purified enzymes exhibit the best affinity for acidic substrates such as casein and can use GTP as well as ATP as phosphoryl donor. They can thus be classified as casein kinase of the G type as previously proposed (Cochet C. et al., (1980) Endocrinology 106, 750-757). Whereas the two moieties could be separated using their different affinities toward a phosphocellulose resin, both purified enzymes appeared indistinguishable on the basis of several molecular and catalytic properties. Both G type casein kinase moieties have an identical sedimentation behavior (5.5 S in the presence of 0.5 M NaCl), yield similar patterns upon electrophoresis under denaturing conditions with three major protein components (42 000, 38 000 and 27 000), and show an ability to undergo self-phosphorylation mostly on the 27 000 component. Both enzymes have the same protein and nucleotide (ATP and GTP) substrate specificity, show similar increases in activity in the presence of polyamines and Mg2+ (optimum at 50 mM) and similar inhibition by NaCl above 0.2 M. The only difference between the two forms of casein kinase (i.e., affinity for phosphocellulose) could not be explained by a different degree of self-phosphorylation or by a limited proteolytic process during handling and purification. These results suggest that the two active moieties may represent isoenzymatic forms of the G type casein kinase activity in bovine adrenal cortex cytosol.
D000255:Adenosine Triphosphate; D000302:Adrenal Cortex / Q000201:enzymology*; D000818:Animals; D047388:Casein Kinases; D002417:Cattle; D002482:Cellulose / Q000031:analogs & derivatives; D002846:Chromatography, Affinity; D006160:Guanosine Triphosphate; D007527:Isoenzymes / Q000302:isolation & purification*; D009712:Nucleotides, Cyclic / Q000494:pharmacology; D010766:Phosphorylation; D011494:Protein Kinases / Q000302:isolation & purification*; D013379:Substrate Specificity
D007527:Isoenzymes; D009712:Nucleotides, Cyclic; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D002482:Cellulose; C008881:phosphocellulose; D011494:Protein Kinases; D047388:Casein Kinases