The subcellular distribution of triphosphoinositide phosphomonoesterase in guinea-pig brain.
1. Some properties of the triphosphoinositide phosphomonoesterase from the homogenates of guinea-pig brain were studied. The enzyme has an optimum pH range 6.7-7.3, is stimulated with KCl at a concentration of 0.1m, and under these conditions has K(m)1.43x10(-4)m. 2. A factor from the ;pH5 supernatant' of guinea-pig brain stimulates the enzyme activity over and above the stimulation produced by KCl. Subcellular fractions of guinea-pig brain varied in their response to the ;pH5 supernatant'. Maximum stimulation was observed with the P(1) fraction, containing myelin and nuclei. 3. An assay system for the enzyme was developed that contained optimum concentrations of both KCl and the ;pH5 supernatant'. Acid phosphatases were inhibited by NaF, but, in contrast with previous work, no EDTA was added to the assay system to inhibit the alkaline phosphatases. This reagent inhibited the triphosphoinositide phosphomonoesterase. It was estimated that the remaining fraction of non-specific phosphatases can account for only 14% of the observed triphosphoinositide phosphomonoesterase activity. 4. Subcellular fractions of guinea-pig brain were characterized by electron microscopy and subcellular markers. The triphosphoinositide phosphomonoesterase exhibited a distribution between the fractions similar to that of 5'-nucleotidase, but different from that of alkaline phosphatase.
D000135:Acid Phosphatase / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002467:Cell Nucleus / Q000201:enzymology; D004492:Edetic Acid; D005459:Fluorides; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D009708:Nucleotidases / Q000378:metabolism; D010716:Phosphatidylinositols; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D013347:Subcellular Fractions
D010716:Phosphatidylinositols; D011506:Proteins; D012313:RNA; D011189:Potassium Chloride; D004492:Edetic Acid; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D010744:Phosphoric Monoester Hydrolases; D005459:Fluorides