High tyrosine-specific protein kinase activity in normal human peripheral blood lymphocytes.
Tyrosine-specific protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) activity was measured in normal human nonadherent peripheral blood lymphocytes using synthetic peptide substrates having sequence homologies with either pp60src or c-myc. A high level of tyrosine-specific protein kinase activity was found associated with the cell particulate fraction (100 000 X g pellet). High-pressure liquid chromatography and phosphoamino acid analysis of the synthetic peptide substrates substantiated the phosphorylation of tyrosine residues by the particulate fraction enzyme. The human enzyme was also capable of phosphorylating a synthetic random polymer of 80% glutamic acid and 20% tyrosine. Enzyme activity was half-maximal with 22 microM Mg X ATP and had apparent Km values for the synthetic peptides from 1.9 to 7.1 mM. The enzyme preferred Mg2+ to Mn2+ for optimal activity and was stimulated 2-5-fold by low levels (0.05%) of some ionic as well as non-ionic detergents including deoxycholate, Nonidet P-40 and Triton X-100. The enzyme activity was not stimulated by N6;O2'-dibutyryl cyclic AMP (100 microM), N6;O2'-dibutyryl cyclic GMP (100 microM), Ca2+ (200 microM), insulin (1 microgram/ml) or homogeneous human T-cell growth factor (3 micrograms/ml) under the conditions used. Alkaline-resistant phosphorylation of particulate proteins in vitro revealed protein bands with Mr 59 000 and 54 000 suggesting that there are endogenous substrates for the human lymphocyte tyrosine protein kinase.
D001798:Blood Proteins / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D010455:Peptides; D010766:Phosphorylation; D011494:Protein Kinases / Q000097:blood*; D011505:Protein-Tyrosine Kinases; D013379:Substrate Specificity
D001798:Blood Proteins; D006133:Growth Substances; D010455:Peptides; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases