title: Simple and unique purification by size-exclusion chromatography for an oligomeric enzyme, rat liver cytosolic acetyl-coenzyme A hydrolase.
abstract: An overview of the purification of an oligomeric enzyme, an extramitochondrial acetyl-coenzyme A hydrolase from rat liver, is presented. The enzyme has been purified to homogeneity using two successive size-exclusion chromatography runs, first for the monomeric and second for the oligomeric form of the enzyme. The sequential gel-filtration steps efficiently removed the contaminants of any molecular size, first of different size from that of the monomeric form of the enzyme (K(av)=0.47 on Superdex 200) and second of different size from that of the oligomeric form (K(av)=0.33), allowing us to purify the enzyme in high purity. This strategy provides an excellent model for purifying many other oligomeric proteins including key enzymes or allosteric enzymes regulating metabolism.
authors.affiliation: Department of Biochemistry, St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki, 216-8511, Kanagawa, Japan. n2sue@marianna-u.ac.jp
authors.forename: Naoya
authors.initials: N
authors.lastname: Suematsu
authors.forename: Kazuki
authors.initials: K
authors.lastname: Okamoto
authors.forename: Fumihide
authors.initials: F
authors.lastname: Isohashi
mesh_terms: D000104:Acetyl-CoA Hydrolase / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D001704:Biopolymers; D002850:Chromatography, Gel / Q000379:methods*; D003600:Cytosol / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D051381:Rats
chemical_list: D001704:Biopolymers; D000104:Acetyl-CoA Hydrolase