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-α-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-α-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.
THE SYNTHESIS AND TURNOVER OF RAT LIVER PEROXISOMES : I. Fractionation of Peroxisome Proteins
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Federico Leighton, Brian Poole, Paul B. Lazarow, Christian De Duve; THE SYNTHESIS AND TURNOVER OF RAT LIVER PEROXISOMES : I. Fractionation of Peroxisome Proteins . J Cell Biol 1 May 1969; 41 (2): 521–535. doi: https://doi.org/10.1083/jcb.41.2.521
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