title: A cytosolic cyclic AMP-dependent protein kinase in Dictyostelium discoideum. I. Properties.
abstract: A cAMP-dependent protein kinase was isolated and partially purified from Dictyostelium discoideum. The cytosolic holoenzyme has an apparent Mr = 160,000-180,000; its activity was stimulated significantly by cAMP when Kemptide served as substrate. The enzyme was dissociated and the regulatory subunit purified by affinity chromatography on 8-aminoethylamino-cAMP. Only one type of regulatory subunit was found; it has an apparent Mr = 41,000 and is a substrate for the in vitro phosphorylation by the homologous catalytic subunit and by purified bovine catalytic subunit. Antibody against the regulatory subunit was prepared. The D. discoideum catalytic subunit was separated from cAMP-independent protein kinase by chromatofocusing. The apparent molecular weight of the catalytic subunit of the D. discoideum cAMP-dependent protein kinase is 33,000 and its pI is 6.4. The enzyme catalyzed the phosphorylation of bovine RII but not of RI regulatory subunit and was inhibited by high concentrations of the inhibitor of mammalian cAMP-dependent protein kinase. The evolution of the functional domains of cAMP-dependent protein kinases is discussed on the basis of a comparison of the analogous D. discoideum and vertebrate enzymes.
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology; D002417:Cattle; D003600:Cytosol / Q000201:enzymology*; D004023:Dictyostelium / Q000201:enzymology*; D047908:Intracellular Signaling Peptides and Proteins; D007526:Isoelectric Point; D046911:Macromolecular Substances; D008970:Molecular Weight; D011494:Protein Kinases / Q000378:metabolism*; D011817:Rabbits
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; C015482:protein kinase modulator; D011494:Protein Kinases