The synthesis and turnover of rat liver peroxisomes. I. Fractionation of peroxisome proteins.
Rat liver peroxisomes isolated by density gradient centrifugation were disrupted at pH 9, and subdivided into a soluble fraction containing 90% of their total proteins and virtually all of their catalase, D-amino acid oxidase, L-alpha-hydroxy acid oxidase and isocitrate dehydrogenase activities, and a core fraction containing urate oxidase and 10% of the total proteins. The soluble proteins were chromatographed on Sephadex G-200, diethylaminoethyl (DEAE)-cellulose, hydroxylapatite, and sulfoethyl (SE)-Sephadex. None of these methods provided complete separation of the protein components, but these could be distributed into peaks in which the specific activities of different enzymes were substantially increased. Catalase, D-amino acid oxidase, and L-alpha-hydroxy acid oxidase contribute a maximum of 16, 2, and 4%, respectively, of the protein of the peroxisome. The contribution of isocitrate dehydrogenase could be as much as 25%, but is probably much less. After dissolution of the cores at pH 11 , no separation between their urate oxidase activity and their protein was achieved by Sephadex G-200 chromatography.
D000429:Alcohol Oxidoreductases / Q000032:analysis; D000818:Animals; D002021:Buffers; D002374:Catalase / Q000032:analysis*; D002498:Centrifugation; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D003594:Cytoplasmic Granules / Q000032:analysis*; D003605:D-Amino-Acid Oxidase / Q000032:analysis; D003576:Electron Transport Complex IV / Q000032:analysis; D005260:Female; D005952:Glucose-6-Phosphatase / Q000032:analysis; D007521:Isocitrate Dehydrogenase / Q000032:analysis; D008099:Liver / Q000166:cytology*; D009940:Organoids / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D051381:Rats; D014503:Urate Oxidase / Q000032:analysis
D002021:Buffers; D011506:Proteins; D000429:Alcohol Oxidoreductases; D007521:Isocitrate Dehydrogenase; D002374:Catalase; D003605:D-Amino-Acid Oxidase; D014503:Urate Oxidase; D003576:Electron Transport Complex IV; D005952:Glucose-6-Phosphatase