Partial purification of ethanolaminephosphotransferase from rat brain microsomes.
Rat brain ethanolaminephosphotransferase (CDPethanolamine : 1,2-diacylglycerol ethanolaminephosphotransferase, EC 2.7.8.1) was solubilized by treating rat brain microsomes with buffered solutions containing octyl glucoside or Triton X-100. The solubilized enzyme was stable both at 4 degrees C and at -18 degrees C. A partial purification was obtained using an ion-exchange chromatographic procedure. The partially purified enzyme showed four major bands in SDS-polyacrylamide gel electrophoresis; its specific activity was increased by a factor of 37 compared to that of the membrane-bound enzyme. Glycerol and diacylglycerol were effective as stabilizers. Phosphatidylcholine, lysophosphatidylcholine and phosphatidylserine increased both the specific activity and the stability of the partially purified enzyme.
D000818:Animals; D001921:Brain / Q000201:enzymology / Q000648:ultrastructure*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004982:Ethanolaminephosphotransferase / Q000302:isolation & purification*; D005960:Glucosides; D005990:Glycerol / Q000494:pharmacology; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology*; D017830:Octoxynol; D010743:Phospholipids / Q000494:pharmacology; D010770:Phosphotransferases / Q000302:isolation & purification*; D011092:Polyethylene Glycols; D051381:Rats; D012995:Solubility
D005960:Glucosides; D010743:Phospholipids; C018619:octyl-beta-D-glucoside; D011092:Polyethylene Glycols; D017830:Octoxynol; D010770:Phosphotransferases; D004982:Ethanolaminephosphotransferase; D005990:Glycerol