Identification of a phosphodiesterase that converts inositol cyclic 1:2-phosphate to inositol 2-phosphate.
Inositol 2-phosphate (Ins(2)P) has been identified in several cell types. The cellular levels of Ins(2)P appear to be directly correlated with the levels of inositol 1:2-cyclic phosphate (cIns(1:2)P) (Ross, T. S., Wang, F. P., and Majerus, P. W. (1992) J. Biol. Chem. 267, 19919-19923). In this study we have detected an enzyme in extracts from CV-1 cells and rat cerebellum that converts cIns(1:2)P to Ins(2)P and inositol 1-phosphate. This enzyme (designated cyclic hydrolase II) is not the same protein previously designated cIns(1:2)P 2-phosphohydrolase (cyclic hydrolase I). The products, heat inactivation curves, pH optima, and metal dependence of these two activities are different, and the two activities were separated by DEAE and gel filtration chromatography. Mixing of cyclic hydrolase I with cyclic hydrolase II does not effect the activity of either. The Km of the CV-1 cyclic hydrolase II for D-cIns(1:2)P is 10 microM. The enzyme is approximately 55 kDa as estimated by gel filtration analysis in the presence of sodium chloride and 120 kDa in its absence.
Ross
T S
TS
Division of Hematology/Oncology, Washington University, St. Louis, Missouri 63110-1093.
Majerus
P W
PW
D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000201:enzymology; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004795:Enzyme Stability; D006358:Hot Temperature; D007295:Inositol Phosphates / Q000378:metabolism*; D010726:Phosphodiesterase Inhibitors / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity
D007295:Inositol Phosphates; D010726:Phosphodiesterase Inhibitors; C010199:inositol cyclic phosphate; C047872:inositol 2-monophosphate; D010727:Phosphoric Diester Hydrolases; C077289:inositol cyclic 1,2-phosphate 1-phosphodiesterase