A membrane-bound activity catalysing phosphatidylinositol breakdown to 1,2-diacylglycerol, D-myoinositol 1:2-cyclic phosphate an D-myoinositol 1-phosphate. Properties and subcellular distribution in rat cerebral cortex.
1. Breakdown of phosphatidylinositol was studied in homogenates and subcellular fractions of rat cerebral cortex by using both membrane-bound and externally added [(32)P]phosphatidylinositol as substrate. 2. In the presence of deoxycholate breakdown followed first-order kinetics at low substrate concentrations ([unk]1mm) and zero-order kinetics at higher concentrations (6-9mm). 3. Maximum breakdown by cerebral-cortex homogenates was approximately 0.5mumol/h per mg of protein and occurred at pH7.0 in the presence of 8mm-phosphatidylinositol, 2mm-CaCl(2) and 2mg of deoxycholate/ml. Activity was abolished by 1mm-ethanedioxybis(ethylamine)tetra-acetate. 4. The products of phosphatidylinositol breakdown were 1,2-diacylglycerol and a mixture of d-myoinositol 1:2-cyclic phosphate (55%) and d-myoinositol 1-phosphate (45%). The two phosphate esters appeared to be produced together and in constant proportions. 5. Some 51% of the activity was particle-bound, with the highest activities in small nerve endings, microsomal material and two synaptic membrane fractions (fractions Mic(20), Mic(100), M(1) 1.0 and M(1) 0.9 respectively), all of which were also rich in acetylcholinesterase and which have been shown to be rich in other surface-membrane enzymes. Much of the particle-bound activity therefore appears to be present in cerebral-cortex plasma membranes. 6. The results are discussed in relation to previously described soluble activities that catalyse the same reaction, and to a possible role of the membrane-bound enzyme in enhanced phosphatidylinositol turnover in externally stimulated cells.
D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002384:Catalysis; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D003840:Deoxycholic Acid / Q000494:pharmacology; D005989:Glycerides / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007294:Inositol / Q000378:metabolism*; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D009186:Myelin Sheath / Q000201:enzymology; D009411:Nerve Endings / Q000201:enzymology; D009994:Osmolar Concentration; D010716:Phosphatidylinositols / Q000096:biosynthesis / Q000378:metabolism*; D010756:Phosphoric Acids / Q000378:metabolism; D010759:Phosphorus Isotopes; D051381:Rats; D013237:Stereoisomerism; D013347:Subcellular Fractions / Q000201:enzymology*; D013570:Synaptic Membranes / Q000201:enzymology
D005989:Glycerides; D010716:Phosphatidylinositols; D010756:Phosphoric Acids; D010759:Phosphorus Isotopes; D003840:Deoxycholic Acid; D007294:Inositol; D000110:Acetylcholinesterase; D002118:Calcium