Histone phosphatase and cyclic nucleotide-stimulated protein kinase from human lymphocytes.
1. Extracts of human peripheral blood lymphocytes contained a histone phosphatase that catalysed the release of P(i) from phosphorylated whole thymus histone. 2. Stimulation of the phosphatase was obtained by concentrations of KCl and NaCl of up to 75mm, and by MgCl(2); CaCl(2) inhibited the enzymic activity. 3. In the absence of MgCl(2), phosphoenol-pyruvate inhibited histone phosphatase activity; this inhibition could be partially reversed by adding MgCl(2) to assays. 4. Lymphocyte extracts contained a protein kinase activity which was maximally stimulated by 1mum-cyclic AMP (adenosine 3':5'-cyclic monophosphate) or by 0.1mm-cyclic GMP (guanosine 3':5'-cyclic monophosphate). 5. Incubation of the enzyme with histone in the absence of ATP or MgCl(2) resulted in the dissociation of the enzyme into a lower-molecular-weight species that was not stimulated by cyclic AMP. This effect could be prevented if ATP and MgCl(2) were present in reaction mixtures before histone and enzyme were allowed to interact. 6. Cyclic AMP also dissociated the kinase into a lower-molecular-weight species. 7. In the presence of 1mum-AMP, half-maximal activities were obtained with 0.92mm-MgCl(2), 6.0mum-ATP and 0.23mg of whole thymus histone/ml.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D001792:Blood Platelets; D002122:Calcium Chloride / Q000494:pharmacology; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D000242:Cyclic AMP / Q000494:pharmacology; D003956:Dialysis; D006657:Histones; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D010759:Phosphorus Isotopes; D010770:Phosphotransferases / Q000302:isolation & purification*; D011188:Potassium / Q000494:pharmacology; D011494:Protein Kinases / Q000302:isolation & purification; D012964:Sodium / Q000494:pharmacology; D012995:Solubility
D006657:Histones; D010759:Phosphorus Isotopes; D010728:Phosphoenolpyruvate; D000255:Adenosine Triphosphate; D012964:Sodium; D000242:Cyclic AMP; D010770:Phosphotransferases; D011494:Protein Kinases; D010744:Phosphoric Monoester Hydrolases; D008274:Magnesium; D002122:Calcium Chloride; D011188:Potassium