Purification and Properties of myo-Inositol-1-Phosphatase from Rat Brain

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Abstract
myo-Inositol-1-phosphatase [EC 3.1.3.25] was purified from a cytosolic fraction of rat brain. The purified enzyme appeared homogeneous on SDS-polyacrylamide gel electrophoresis and its molecular weight was estimated to be 29,000. The molecular weight of the native enzyme was 55,000 as determined by molecular sieve chromatography. These values indicated that the native enzyme was composed of two identical subunits. The isoelectric point of the enzyme was 4.6. The enzyme hydrolyzed inositol-1-phosphate, 2′-AMP, 2′-AMP, β-glycerophosphate, and α-glycerophosphate; the ratio of the reaction rates was 100: 84 : 73 : 64 : 32. The Km values for inositol-1-phosphate, 2′-AMP, and β-glycerophosphate were 1.2 × 10−4 M, 1.9 × 10−4 M, and 7.7 × 10−4 M, respectively. Mn2+ and Ca2+ were strong competitive inhibitors against Mg2+, with K1 values of 3 μM and 20 μM, respectively. This result suggests that myo-inositol-1-phosphatase might be regulated by intracellular Ca2+ and/or Mn2+. Li+, which is known to show a therapeutic effect on manic-depressive disease and also to prolong the intrinsic periods of circadian rhythms in various organisms, was a potent uncompetitive inhibitor and inhibited 50% of the activity at 1 mM. The possibility that myo-inositol-1-phosphatase and inositol phospholipid metabolism are involved in circadian rhythm oscillation is discussed in terms of Li actions.