Simple and unique purification by size-exclusion chromatography for an oligomeric enzyme, rat liver cytosolic acetyl-coenzyme A hydrolase.
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.
Suematsu
Naoya
N
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
Okamoto
Kazuki
K
Isohashi
Fumihide
F
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
D001704:Biopolymers; D000104:Acetyl-CoA Hydrolase