Isolation and characterization of peroxisomes from the renal cortex of beef, sheep, and cat.
The isolation and characterization of highly purified and structurally well-preserved peroxisomes from the renal cortex of different mammalian species (beef, sheep, and cat) is reported. Renal cortex tissue was homogenized and a peroxisome-enriched light mitochondrial fraction was prepared by differential centrifugation. This was subfractionated by density-dependent banding on a linear gradient of metrizamide (1.12-1.26 g/cm3) using a Beckman VTi 50 vertical rotor. Peroxisomes banded at a mean density of 1.225 cm3. Ultrastructural morphometric examination revealed that peroxisomes made up 97 to 98% of the isolated fractions. By biochemical analysis the contamination with marker enzymes of mitochondria and lysosomes was extremely low. The specific activity of catalase was enriched, depending on the species, between 28- and 38-fold over the homogenate. Peroxisome preparations from all three species exhibited a high but varying level of activity for cyanide-insensitive lipid beta-oxidation. In beef and sheep preparations a small amount of esterase activity cosediments with peroxisomes. These peroxisomes show distinct structural membrane associations with smooth elements of ER. Urate oxidase, a marker enzyme for rat liver peroxisomes, is found only in peroxisomes prepared from beef kidney cortex, with sheep and cat preparations being negative. This correlated with the occurrence of polytubular inclusions in the beef kidney peroxisomes. The large size and the angular shape of isolated peroxisomes as well as the presence of paracrystalline matrical inclusions imply that the majority of peroxisomes are derived from the epithelial cells of the proximal tubule of the kidney cortex. The significant differences found in the characteristics of the renal peroxisomes in three different species investigated, demonstrate the remarkable adaptability and plasticity of this organelle.
D000818:Animals; D002374:Catalase / Q000032:analysis; D002415:Cats; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D003576:Electron Transport Complex IV / Q000032:analysis; D005966:Glucuronidase / Q000032:analysis; D007672:Kidney Cortex / Q000648:ultrastructure*; D008793:Metrizamide; D008830:Microbodies / Q000032:analysis / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure*; D008854:Microscopy, Electron; D010316:Particle Size; D012756:Sheep; D013347:Subcellular Fractions / Q000032:analysis
D002374:Catalase; D003576:Electron Transport Complex IV; D005966:Glucuronidase; D008793:Metrizamide