Identification of glycosylphosphatidylinositol-specific phospholipases C in mouse brain membranes.
Using the membrane form of variant surface glycoprotein from Trypanosoma equiperdum labelled with [3H]myristate as a substrate, we identified two glycosylphosphatidylinositol phospholipase C enzymic activities in mouse brain. These activities were associated with particulate membrane fractions. They were characterized by their pH activity maxima and sensitivity to activators and ion chelators. One of the activities was maximal at acidic pH, stimulated by butanol, sensitive to cation chelator and insensitive to manganese. The activity of the other was maximal at neutral pH, stimulated by the detergent deoxycholate and independent of the presence of cation chelator or calcium. On membrane subfractionation, the acidic butanol-stimulated activity was found mainly associated with the lysosomal compartment, whereas the neutral deoxycholate-stimulated activity sediments with the myelin and plasma membrane compartment. These activities could be differentiated from particulate phosphatidylinositol phospholipases C, whose acidic lysosomal form is sensitive to manganese and insensitive to cation chelator or butanol, whereas the deoxycholate-activated enzymes are Ca2(+)-dependent.
Fouchier
F
F
Institut de Chimie Biologique, CNRS URA 202, Université d'Aix-Marseille, France.
Baltz
T
T
Rougon
G
G
D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D000440:Butanols / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D002614:Chelating Agents / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006017:Glycolipids / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000201:enzymology; D008345:Manganese / Q000494:pharmacology; D051379:Mice; D009186:Myelin Sheath / Q000201:enzymology; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010716:Phosphatidylinositols / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D010738:Type C Phospholipases / Q000032:analysis* / Q000378:metabolism; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000378:metabolism
D000440:Butanols; D002614:Chelating Agents; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D010716:Phosphatidylinositols; D014643:Variant Surface Glycoproteins, Trypanosoma; D003840:Deoxycholic Acid; D008345:Manganese; D010727:Phosphoric Diester Hydrolases; D010738:Type C Phospholipases; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D002118:Calcium