Endogenous phosphorylation of microsomal proteins in bovine corpus luteum. Tenfold activation by adenosine 3':5'-cyclic monophosphate.
Free ribosomes and a smooth-microsomal fraction were prepared from bovine corpus luteum. Both preparations will self-phosphorylate when incubated with Mg(2+) and ATP, but at low concentrations of Mg(2+) and ATP the self-phosphorylation of the smooth-microsomal fraction was much more dependent on cyclic AMP than was that of free ribosomes, stimulation by the nucleotide being up to 10-fold in the former case. The self-phosphorylation of the smooth-microsomal fraction was studied further. The reaction bears similarities to that brought about by soluble cyclic AMP-dependent protein kinase, being inhibited by Ca(2+) and the heat-stable inhibitor protein from skeletal muscle. Cyclic GMP will activate the reaction at concentrations higher than those required for full activation by cyclic AMP. In the presence of cyclic AMP, phosphate bound to protein is found almost exclusively as phosphoserine. Several proteins are phosphorylated, as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, and the phosphorylation of all of them is markedly stimulated by cyclic AMP. If the reaction is carried out at high concentrations of Mg(2+) and ATP, a distinct cyclic AMP-independent phosphorylation is observed. This activity is not inhibited by the heat-stable inhibitor protein, and phosphate is found esterified with both threonine and serine residues.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002458:Cell Fractionation; D003338:Corpus Luteum / Q000378:metabolism*; D000242:Cyclic AMP / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008274:Magnesium / Q000494:pharmacology; D008861:Microsomes / Q000378:metabolism*; D010750:Phosphoproteins / Q000032:analysis; D010758:Phosphorus / Q000378:metabolism*; D047428:Protein Kinase Inhibitors; D012270:Ribosomes / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate
D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D010758:Phosphorus; D012967:Sodium Dodecyl Sulfate; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D008274:Magnesium; D002118:Calcium