Squalene synthetase. Solubilization and partial purification of squalene synthetase, copurification of presqualene pyrophosphate and squalene synthetase activities.
Squalene synthetase (farnesyldiphosphate:farnesyldiphosphate farnesyltransferase, EC 2.5.1.21) is an intrinsic microsomal protein that catalyzes the synthesis of squalene from farnesyl pyrophosphate via the intermediate presqualene pyrophosphate. We have solubilized this enzyme from yeast with a mixture of the detergents N-octyl beta-D-glucopyranoside and Lubrol PX. Approximately 50-fold purification of the solubilized activities has been achieved by chromatography on DEAE-cellulose and hydroxylapatite and by isoelectric focusing. The most highly purified preparation has one major band of protein with a molecular weight of 53,000 as estimated by electrophoresis under denaturing conditions. The enzyme may also have been modified by proteolysis during isolation since a 47,000 molecular weight species was also found. The two activities, presqualene pyrophosphate synthetase and squalene synthetase, copurified during isolation.
D002848:Chromatography, DEAE-Cellulose; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000302:isolation & purification*; D005960:Glucosides; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010088:Oxidoreductases / Q000302:isolation & purification*; D000077423:Polidocanol; D011092:Polyethylene Glycols; D011106:Polyisoprenyl Phosphates / Q000302:isolation & purification* / Q000378:metabolism; D012717:Sesquiterpenes; D012995:Solubility; D013185:Squalene / Q000378:metabolism
D005960:Glucosides; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; D000077423:Polidocanol; C018619:octyl-beta-D-glucoside; C017672:presqualene pyrophosphate; D011092:Polyethylene Glycols; C004808:farnesyl pyrophosphate; D013185:Squalene; D010088:Oxidoreductases; D013186:Farnesyl-Diphosphate Farnesyltransferase