title: Adenosine 3':5'-cyclic monophosphate-dependent and plasma-membrane-associated protein kinase(s) from bovine corpus luteum.
abstract: Plasma-membrane fractions FI and FII isolated from bovine corpus luteum by discontinuous sucrose-density-gradient centrifugation, at sucrose-density interfaces of 1.14/1.16 and 1.16/1.18 respectively, contained membrane-associated protein kinases that phosphorylated both the structural proteins of membranes as well as exogenously added protein substrates. Both fractions were characterized with respect to endogenous and exogenous protein substrate specificity, pH-dependence, effect of bivalent metal ions and sensitivity toward cyclic nucleotides. These membrane-associated kinases showed an optimum pH of 6.0 and had an absolute requirement for bivalent metal ions such as Mg2+, Mn2+, or Co2+ that cannot be replaced by Ca2+. Both the activities were stimulated two- to four-fold by cyclic AMP in vitro with an apparent Km of 83 and 50 nM for fractions FI and FII respectively. Other cyclic 3':5'-nucleotides were effective only at higher concentrations, but even the most effective, cyclic IMP, showed a stimulation nearly an order of magnitude lower than that of cyclic AMP. In contrast, stimulation by cyclic dTMP and cyclic dAMP was very weak. Cyclic AMP showed no significant effect on the apparent Km value of both enzymes for histone and MgCl2 but it somewhat decreased the Km value for ATP. Nucleoside triphosphates like GTP, CTP and UTP inhibited the transfer of [32P]Pi from [gamma-32P]ATP into mixed histone catalysed by membrane-associated kinases either in the presence or in the absence of cyclic AMP. In addition to protein kinases, these membrane fractions also possessed cyclic AMP-binding activities. The apparent association constant (Kalpha) for cyclic AMP binding was 1.0 X 10(10) and 2.6 X 10(10) M for FI and FII membrane fractions respectively.
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002364:Caseins / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D003035:Cobalt / Q000494:pharmacology; D003338:Corpus Luteum / Q000201:enzymology*; D000242:Cyclic AMP / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D005260:Female; D006062:Gonadotropins / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009705:Nucleosides / Q000378:metabolism; D009712:Nucleotides, Cyclic / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011479:Protamines / Q000378:metabolism; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*
chemical_list: D002364:Caseins; D006062:Gonadotropins; D006657:Histones; D009705:Nucleosides; D009712:Nucleotides, Cyclic; D010710:Phosphates; D011479:Protamines; D003035:Cobalt; D008345:Manganese; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D011494:Protein Kinases; D008274:Magnesium; D002118:Calcium