Comparison of the activities of a multiple inositol polyphosphate phosphatase obtained from several sources: a search for heterogeneity in this enzyme.
A multiple inositol polyphosphate phosphatase (formerly known as inositol 1,3,4,5-tetrakisphosphate 3-phosphatase) was purified approx. 22,000-fold from rat liver. The final preparation migrated on SDS/PAGE as a doublet with a mean apparent molecular mass of 47 kDa. Upon size-exclusion chromatography, the enzyme was eluted with an apparent molecular mass of 36 kDa. This enzyme was approximately evenly distributed between the 'rough' and 'smooth' subfractions of endoplasmic reticulum. There was a 20-fold range of specific activities of this phosphatase in CHAPS-solubilized particulate fractions prepared from the following rat tissues: liver, heart, kidney, testis and brain. However, each of these extracts contained different amounts of endogenous inhibitors of enzyme activity. After removal of these inhibitors by MonoQ anion-exchange chromatography, there was only a 2.5-fold range of specific activities; kidney contained the most and brain contained the least. We prepared and characterized polyclonal antiserum to the hepatic phosphatase, which immunoprecipitated 85-100% of both particulate and soluble phosphatase activities. The antiserum also immunoprecipitated, with equivalent efficacy, CHAPS-solubilized phosphatase activities from heart, kidney, testis, brain and erythrocytes (all prepared from rat). Our data strengthen the case that the function of the mammalian phosphatase is unrelated to the metabolism of Ca(2+)-mobilizing cellular signals. The CHAPS-solubilized phosphatase from turkey erythrocytes was not immunoprecipitated by the polyclonal antiserum, and is therefore an isoform that is structurally distinct, and possibly functionally unique.
Inositol Lipid Section, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.
A
A
Craxton
N
N
Ali
S B
SB
Shears
D000818:Animals; D001921:Brain / Q000201:enzymology; D002451:Cell Compartmentation; D003429:Cross Reactions; D004721:Endoplasmic Reticulum / Q000201:enzymology; D004789:Enzyme Activation; D014644:Genetic Variation; D007295:Inositol Phosphates / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008297:Male; D008566:Membranes / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D010744:Phosphoric Monoester Hydrolases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013737:Testis / Q000201:enzymology; D014018:Tissue Distribution
D007295:Inositol Phosphates; C081655:inositol-1,3,4,5-tetrakisphosphate; D010744:Phosphoric Monoester Hydrolases; C064119:multiple inositol-polyphosphate phosphatase