[A nonisotopic enzymolysis assay method of phosphatidylinositol kinase].
A nonisotopic enzymolysis assay method of phosphatidylinositol kinase (PIK) has been developed. Phosphatidylinositol 4-phosphate (PIP), phosphorylated from phosphatidylinositol (PI) by PIK was hydrolyzed with phosphainositide-specific phospholipase C. The product, inositol 1, 4-bisphosphate (IP2), was separated from inositol 1-phosphate (IP) with Dowex-1 column chromatography. Then, the IP2 was hydrolyzed by alkaline phosphatase to yield PI, and the PI was determined by colorimetry. The PIK activity was defined as PI nmol/mg protein per min. The recovery was 91%, CV = 6.5%
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Liu
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Ma
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D019870:1-Phosphatidylinositol 4-Kinase; D002845:Chromatography / Q000379:methods; D006801:Humans; D010770:Phosphotransferases / Q000378:metabolism*
D010770:Phosphotransferases; D019870:1-Phosphatidylinositol 4-Kinase