Protein kinases in normal human blood cells.
Protein kinases active on basic and acidic artificial substrates were investigated in normal human erythrocytes, platelets, polymorphonuclear and mononuclear cells. These two types of protein kinases were partially purified by affinity chromatography, then assayed for their enzymatic activity using [gamma-32P]ATP or GTP as phosphoryl donor. Partially purified kinases active on acidic substrates were subjected to high-performance liquid chromatography (HPLC). Protein kinases active on basic substrates were analyzed by cellulose acetate electrophoresis of crude cellular extracts and the influence of 3'5' cyclic AMP was studied. Three forms of casein-phosvitin kinases could be distinguished according to their molecular weight (165 K, 38 K, and 31 K). The 165 K species, in contrast to the light species, can use GTP instead of ATP as phosphoryl donor and corresponds to the "so-called" casein kinase 2. This form is very sensitive to proteolysis and, when partial purification is performed without the addition of various antiproteolytic agents, it is degraded into 120-135 K and 105-115 K active species; this artefactual degradative process is especially active in platelet extracts. As many as eight different active bands of histone and protamine kinases can be separated by cellulose acetate electrophoresis, several of them being stimulated by cyclic AMP. Isozymic patterns of protein kinases, levels of activity on the different substrates, and utilization of ATP and GTP were found to be specific for each cell type. These results suggest the possibility of using protein kinases as markers for cell differentiation.
D001773:Blood Cells / Q000201:enzymology*; D001792:Blood Platelets / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D004588:Electrophoresis, Cellulose Acetate; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology; D009504:Neutrophils / Q000201:enzymology; D011494:Protein Kinases / Q000097:blood* / Q000302:isolation & purification
D011494:Protein Kinases