Purification of a Phosphatidylinositol 4-Phosphate Kinase from Bovine Brain Membranes |
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Authors: | Albrecht Moritz Pierre N E De GraanPeter F Ekhart Willem H Gispen Karel W A Wirtz |
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Institution: | Center for Biomembranes and Lipid Enzymology, University of Utrecht, The Netherlands. |
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Abstract: | A phosphatidylinositol 4-phosphate (PIP) kinase (EC 2.7.1.68) was purified from bovine brain membranes in a six-step procedure involving solubilization of the enzyme with 170 mM NaCl followed by chromatography on diethylaminoethyl-cellulose, phosphocellulose, Ultrogel AcA44, hydroxylapatite, and ATP-agarose. The enzyme preparation was nearly homogeneous and was purified 5,600-fold with a final specific activity of 85 nmol/min/mg of protein and a yield of 20%. Its molecular mass was 110 kilodaltons, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was specific for PIP; phosphorylation of phosphatidylinositol and diacylglycerol was not observed. |
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Keywords: | Phosphatidylinositol 4-phosphate kinase Enzyme purification |
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