Open AccessPurification and characterization of bovine brain type I phosphatidylinositol kinase
Author(s) -
MORGAN Sarah J.,
SMITH Anthony D.,
PARKER Peter J.
Publication year - 1990
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1990.tb19185.x
Subject(s) - phosphatidylinositol , kinase , biochemistry , enzyme , chemistry , substrate (aquarium) , polyacrylamide gel electrophoresis , gel electrophoresis , microbiology and biotechnology , biology , ecology
Investigation into the phosphatidylinositol kinase activities in bovine brain has revealed the presence of a type I PtdIns kinase activity. This classification is based upon potent inhibition by neutral detergent and the production of a phosphatidylinositol phosphate that can be distinguished from phosphatidyl‐inositol 4‐phosphate [PtdIns(4) P ] by thin‐layer chromatography. The enzyme has been substantially purified and the activity is associated with an 85‐kDa polypeptide on SDS/polyacrylamide gel electrophoresis. Analysis of the product confirms the identification of the enzyme as a type I PtdIns kinase. The purified kinase has been characterized with respect to substrate dependence (Mg 2+ , ATP, PtdIns), substrate presentation (pure lipid versus mixed micelle) and specificity [PtdIns versus PtdIns(4) P and phosphatidylinositol 4,5‐bisphosphate].