title: Characterization and partial purification of squalene-2,3-oxide cyclase from Saccharomyces cerevisiae.
abstract: The membrane nature of squalene oxide cyclase from Saccharomyces cerevisiae was investigated by comparing properties of the enzyme recovered from both microsomes and the soluble fraction of the yeast homogenate. The "apparent soluble" form and microsomal form of the enzyme were both stimulated by the presence of mammalian soluble cytoplasm and corresponded to one another in response to detergents Triton X-100 and Triton X-114. The observed strong dependence of the enzyme activity on the presence of detergents and the behavior of the enzyme after Triton X-114 phase separation were peculiar to a lipophilic membrane-bound enzyme. A study of the conditions required to extract the enzyme from microsomes confirmed the lipophilic character of the enzyme. Microsomes, exposed to ipotonic conditions to remove peripheral membrane proteins, retained most of the enzyme activity within the integral protein fraction. Quantitative dissociation of the enzyme from membranes occurred only if microsomes were treated with detergents (Triton X-100 or octylglucoside) at concentrations which alter membrane integrity. The squalene oxide cyclase was purified 140 times from yeast microsomes by (a) removal of peripheral proteins, (b) extraction of the enzyme from the integral protein fraction with octylglucoside, and (c) separation of the solubilized proteins by DEAE Bio-Gel A chromatography. Removal of the peripheral proteins seemed to be a key step necessary for obtaining high yields.
authors.affiliation: Istituto di Chimica Farmaceutica Applicata, Torino, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Balliano
authors.forename: F
authors.initials: F
authors.lastname: Viola
authors.forename: M
authors.initials: M
authors.lastname: Ceruti
authors.forename: L
authors.initials: L
authors.lastname: Cattel
mesh_terms: D003902:Detergents / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D019751:Intramolecular Transferases; D007535:Isomerases / Q000737:chemistry* / Q000302:isolation & purification; D008861:Microsomes / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012995:Solubility; D013696:Temperature
chemical_list: D003902:Detergents; D007535:Isomerases; D019751:Intramolecular Transferases; C021924:lanosterol synthase