Subcellular fractionation by differential and zonal centrifugation of the trypanosomatid Crithidia fasciculata.
Homogenates of Crithidia fasciculata were fractionated by differential centrifugation. Mitochondria were sedimented quantitatively at 10(4) g-min and accounted for approximately 10% of the total recovered protein. Catalase was found exclusively in the supernatant fraction whilst NADH:cytochrome c oxidoreductase and p-nitrophenylphosphatase were found in all the fractions. Zonal centrifugation confirmed that catalase was non-sedimentable. Clean separation of mitochondria was obtained in both high-speed and rate zonal experiments, but no NADH:cytochrome c oxidoreductase activity could be detected in these organelles. Separation of large lysosomal vacuoles which contained p-nitrophenylphosphatase activity was obtained and these were clearly resolved from mitochondria by both high-speed and rate zonal centrifugation.
D002458:Cell Fractionation; D002501:Centrifugation, Zonal; D003579:Cytochrome Reductases / Q000032:analysis; D056890:Eukaryota / Q000032:analysis*; D008291:Malate Dehydrogenase / Q000032:analysis; D013347:Subcellular Fractions / Q000032:analysis; D014345:Trypanosoma / Q000201:enzymology
D008291:Malate Dehydrogenase; D003579:Cytochrome Reductases