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Purification of a Phosphatidylinositol 4‐Phosphate Kinase from Bovine Brain Membranes
Author(s) -
Moritz Albrecht,
Graan Pierre N. E.,
Ekhart Peter F.,
Gispen Willem H.,
Wirtz Karel W. A.
Publication year - 1990
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1990.tb13322.x
Subject(s) - phosphatidylinositol , chemistry , chromatography , biochemistry , enzyme , gel electrophoresis , membrane , sodium dodecyl sulfate , phosphorylation , polyacrylamide gel electrophoresis , agarose , phosphate
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 m M 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.