Phosphomonoesterase hydrolysis of polyphosphoinositides in rat kidney: Properties and subcellular localization of the enzyme system.
Tthe properties of diphosphoinositide and triphosphoinositide phosphatases from rat kidney homogenate were studied in an assay system in which non-specific phosphatase activity was eliminated. The enzymes were not completely metal-ion dependent and were activated by Mg2+. The detergent sodium deoxycholate, Triton X-100 and Cutscum inhibited the reaction; cetyltrimethylammonium bromide only activated when added with the subtrates and in the presence Mg2+. Both enzymes had a pH optimum of 7.5. Ca2+ and Li+ both activated triphosphoinositide phosphatase, but Ca2+ inhibited and L+ had little effect on diphosphoinositide phosphatase. Cyclic AMP had no effect on either enzyme. The enzymes were three times more active in kidney cortex than in the medulla. On subcellular fractionation of kidney-cortex homogenates by differential and density-gradient centrifugation, the distribution of the enzymes resembled that of thiamin pyrophosphatase (assayed in the absence of ATP), suggesting localization in the Golgi complex. However, the distribution differed from that of the liver Golgimarker galactosyltransferase. Activities of both diphosphoinositide and triphosphoinositide phosphatases and thiamin pyrophosphatase were low in purified brush-border fragments. Further experiments indicate that at least part of the phosphatase activity is soluble.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002593:Cetrimonium Compounds / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000494:pharmacology; D005260:Female; D005700:Galactosyltransferases / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology; D007668:Kidney / Q000201:enzymology* / Q000648:ultrastructure; D007672:Kidney Cortex / Q000201:enzymology; D007679:Kidney Medulla / Q000201:enzymology; D008094:Lithium / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D008861:Microsomes / Q000201:enzymology; D000442:Octanols / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats; D013834:Thiamine Pyrophosphatase / Q000378:metabolism
D002593:Cetrimonium Compounds; D000442:Octanols; D010716:Phosphatidylinositols; D003840:Deoxycholic Acid; D011092:Polyethylene Glycols; D008094:Lithium; D000242:Cyclic AMP; D005700:Galactosyltransferases; D010744:Phosphoric Monoester Hydrolases; D013834:Thiamine Pyrophosphatase; D008274:Magnesium; D002118:Calcium