Characterization of protein kinases from bovine parotid glands. The effect of tolbutamide and its derivative on these partially purified enzymes.
1. Four fractions of protein kinase (EC 2.7.1.37) activity (Peak IH, IIH, IIIC and IVC) have been resolved and partially purified from the 100 000 X g supernatant fraction of bovine parotid glands by DEAE-cellulose and phosphocellulose chromatographies. 2. The protein kinases of Peak IH and IIH were adenosine 3',5'-monophosphate (cyclic AMP) -dependent and had similar enzymic properties. The enzyme activities of Peak IIIC and IVC were cyclic-AMP independent, but there were some distinct differences between their properties. The protein kinase in Peak IIIC was activated by 0.2 M NaCl or KCl and phosphorylated casein preferentially as the substrate, utilizing only ATP as a phosphate donor. On the other hand, the protein kinase in Peak IVC was inhibited by univalent salts and preferred phosvitin to casein, utilizing either ATP or GTP as a phosphate donor. 3. Tolbutamide increased the Km value for ATP and the dissociation constant for cyclic AMP, resulting in the inhibition of cyclic-AMP dependent protein kinase activity in the presence of cyclic AMP. Tolbtamide and its carboxy derivative, 1-butyl-3-p-carboxyphenylsulfonylurea, exerted almost no inhibitory effect on either the cyclic-AMP dependent protein kinase activities in the absence of cyclic AMP or on the cyclic-AMP independent protein kinase activities.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D000242:Cyclic AMP / Q000494:pharmacology; D006152:Cyclic GMP / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010306:Parotid Gland / Q000187:drug effects / Q000201:enzymology*; D011189:Potassium Chloride / Q000494:pharmacology; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D014044:Tolbutamide / Q000031:analogs & derivatives* / Q000494:pharmacology*
D012965:Sodium Chloride; D011189:Potassium Chloride; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D014044:Tolbutamide; D000242:Cyclic AMP; D011494:Protein Kinases; D006152:Cyclic GMP