title: Cyclic AMP-dependent protein kinase in ovarian follicle cells of starfish Asterina pectinifera.
abstract: Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein kinase (PKA) in ovarian follicle cells of the starfish Asterina pectinifera was studied. Protein kinase activity in follicle cell homogenate was activated by cAMP in a dose-dependent manner, and Ka was obtained with 10(-7) M cAMP. The PKA activity required Mg2+ at concentrations between 2 and 10 mM. On Sephacryl S-300 column chromatography of partially purified PKA, Mr of the holoenzyme was estimated to be about 180,000. [2,3-3H]cAMP binding activity also suggested a regulatory subunit of Mr about 50,000. DE-52 column chromatography of the cell extract resolved the enzyme activity into two peaks, which eluted between 0.05 and 0.1 M NaCl (type I), and between 0.15 and 0.25 M NaCl (type II). The type I enzyme was the predominant form of PKA in starfish follicle cells. In a cell-free system, a 70 kDa protein was phosphorylated during incubation with [gamma-32P]ATP in the presence of cAMP. These results suggest that PKA stimulates the phosphorylation of a 70 kDa protein following an increase in the level of cAMP.
authors.affiliation: Department of Biology, School of Education, Waseda University, Tokyo, Japan. bio-mita@mn.waseda.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Mita
authors.forename: I
authors.initials: I
authors.lastname: Yasumasu
authors.forename: Y
authors.initials: Y
authors.lastname: Nagahama
mesh_terms: D000818:Animals; D002852:Chromatography, Ion Exchange; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D006080:Ovarian Follicle / Q000201:enzymology*; D010750:Phosphoproteins / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D013215:Starfish / Q000201:enzymology*
chemical_list: D010750:Phosphoproteins; D010761:Phosphorus Radioisotopes; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases