Author:

Keywords:

sterol carrier protein 2/3-oxoacyl-coa thiolase, peroxisomes (rat liver), lipid-transfer protein, coenzyme-a thiolase, amino-acid-sequence, beta-oxidation, 3-oxoacyl-coa thiolase, kinetic mechanism, 58-kda protein, purification, metabolism, Science & Technology, Life Sciences & Biomedicine, Biochemical Research Methods, Biochemistry & Molecular Biology, Biotechnology & Applied Microbiology, sterol carrier protein 2/3-oxoacyl-CoA thiolase, LIPID-TRANSFER PROTEIN, COENZYME-A THIOLASE, AMINO-ACID-SEQUENCE, BETA-OXIDATION, 3-OXOACYL-COA THIOLASE, KINETIC MECHANISM, 58-KDA PROTEIN, PURIFICATION, METABOLISM, Acetyl-CoA C-Acetyltransferase, Animals, Carrier Proteins, Chromatography, Agarose, Chromatography, Gel, Cross-Linking Reagents, Electrophoresis, Polyacrylamide Gel, Immunoblotting, Isoenzymes, Liver, Male, Peroxisomes, Protein Structure, Quaternary, Rabbits, Rats, Rats, Wistar, Sterols, 0601 Biochemistry and Cell Biology, 0699 Other Biological Sciences, 3101 Biochemistry and cell biology

Abstract:

In the present report we describe a method for the complete purification of native sterol carrier protein 2/3-oxoacyl-CoA thiolase (SCP-2/thiolase) from normal rat liver peroxisomes. The isolation procedure is based on the alteration in chromatographic properties of the enzyme in the presence of low concentrations of CoA. The purified preparation of SCP-2/thiolase consisted of 58- and 46-kDa polypeptides. Peroxisomes prepared freshly from normal rat liver contained three SCP-2/thiolase isoforms, separable by conventional chromatography. Immunochemical, molecular sieving, and chemical cross-linking experiments indicated that these isoforms represent thiolytically active homo- and heterodimeric combinations of the 46- and 58-kDa subunits (2 x 58, 58-46, and 2 x 46-kDa proteins). (C) 2000 Academic Press.