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J. Biol. Chem., Vol. 265, Issue 36, 22086-22089, 12, 1990

A novel pathway for the formation of phosphatidylinositol 3,4- bisphosphate. Phosphorylation of phosphatidylinositol 3-monophosphate by phosphatidylinositol-3-monophosphate 4-kinase

K Yamamoto, A Graziani, C Carpenter, LC Cantley and EG Lapetina
Division of Cell Biology, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.

Three polyphosphoinositides containing phosphate at the D-3 position of the inositol ring can be generated in vitro by phosphorylation of phosphatidylinositol, phosphatidylinositol 4-phosphate, and phosphatidylinositol 4,5-bisphosphate by a phosphatidylinositol-3- kinase (Auger, K. R., Serunian L. A., Soltoff, S. P., Libby, P., and Cantley, L. C. (1989) Cell 57, 167-175. An alternative pathway for in vivo synthesis of one of these lipids was recently suggested: phosphatidylinositol 3,4-bisphosphate could be produced by phosphorylation of phosphatidylinositol 3-phosphate at the D-4 position of the inositol ring (Yamamoto, K., and Lapetina, E. G. (1990) Biochem. Biophys. Res. Commun. 168, 466-472). Here we demonstrate the presence of an enzyme in human platelets that phosphorylates [32P]phosphatidylinositol 3-phosphate to produce [32P]phosphatidylinositol 3,4-bisphosphate. This enzyme is Mg2(+)- dependent and its apparent molecular mass is approximately 150 kDa as estimated by sucrose gradient centrifugation and gel filtration chromatography. Unlike the major phosphatidylinositol-4-kinase in platelets that is stimulated by the detergent Nonidet P-40, the phosphatidylinositol-3-phosphate-4-kinase is inhibited by Nonidet P-40. Both activities are also differentiated by the action of adenosine. The discovery of this new enzyme raises the possibility that multiple pathways exists for generating D-3 phosphorylated phosphoinositides.
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