[Properties of tyrosine-specific protein kinase from rat sarcoma cells].
A procedure for measuring the activity of tyrosine-specific protein kinases was developed. The method is based on the fact that the phosphoryl groups of phosphotyrosine residues of phosphorylated protein substrates are not hydrolyzed in alkaline solutions, whereas the phosphoserine and phosphothreonine residues lose their phosphoryl groups upon heating in 2 N KOH. It was demonstrated that rat sarcoma XC cells in culture and in solid tumours contain tyrosine-specific protein kinase (TPK). The TPK is localized in the membrane fraction of the cells and is solubilized by 1% Triton X-100. Mn2+-ATP is a nucleotide substrate for TPK. Among other protein substrates, TPK strongly phosphorylates histones H5 and H2a, weakly phosphorylates histones H2b, H4 and H1 but does not phosphorylate protamine, casein, vinculin or angiotensin II. The optimal concentration of Mn2+ for the reaction is 2 mM; the Km values for ATP and histone H5 are 2-3 microM and 2.5 mg/ml, respectively. Tyrosine-specific protein kinases phosphorylating histone H5 were detected in the membrane fraction isolated from different mammalian tissues, e. g., spleen, heart, liver, brain and lungs, as well as from human intestinal mucosa and mucosal adenocarcinoma. These results suggest that tyrosine-specific protein kinases phosphorylating histone H5 are present in a vast majority of mammalian tissues.
Petukhov
S P
SP
Lebedeva
E L
EL
Bulargina
T V
TV
Severin
E S
ES
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001358:Avian Sarcoma Viruses; D001667:Binding, Competitive; D002461:Cell Line, Transformed; D006657:Histones / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012513:Sarcoma, Experimental / Q000201:enzymology*; D013045:Species Specificity; D013379:Substrate Specificity
D006657:Histones; D008565:Membrane Proteins; D000255:Adenosine Triphosphate; D011505:Protein-Tyrosine Kinases
Izuchenie svoĭstv tirozinspetsificheskoĭ proteinkinazy iz kletok sarkomy krysy.