Purification and characterization of phosphatidylinositol 4-phosphate 5-kinases.
We detail the purification and characterization of three distinct isoforms of PtdIns4P 5-kinase present in bovine brain. One of these, PtdIns4P 5-kinase C, was purified to apparent homogeneity, and SDS/PAGE analysis demonstrated a single polypeptide and molecular mass 53 KDa. These three isoforms were shown to differ in their kinetic properties, and immunological characterization with an antibody raised to PtdIns4P 5-kinase C demonstrated that this isoform was unrelated to the other two. Furthermore, PtdIns4P 5-kinase C was shown to be the bovine brain homologue of the Type II PtdIns4P 5-kinase previously purified from human erythrocytes [Bazenet, Ruano, Brockman & Anderson (1990) J. Biol. Chem. 265, 18012-18022].
Divecha
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N
AFRC Institute of Animal Physiology and Genetics Research, Cambridge, U.K.
Brooksbank
C E
CE
Irvine
R F
RF
D000818:Animals; D001921:Brain / Q000201:enzymology; D002417:Cattle; D003429:Cross Reactions; D003600:Cytosol / Q000201:enzymology; D004912:Erythrocytes / Q000276:immunology; D006801:Humans; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification; D007700:Kinetics; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000378:metabolism; D010770:Phosphotransferases / Q000276:immunology / Q000302:isolation & purification*; D017853:Phosphotransferases (Alcohol Group Acceptor); D013379:Substrate Specificity
D007527:Isoenzymes; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; C037178:phosphatidylinositol 4-phosphate; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C020077:1-phosphatidylinositol-4-phosphate 5-kinase