SUMMARY: Homogenates from were fractionated by differential centrifugation. All the mitochondria (located by measurement of cytochrome oxidase) and most of the peroxisomes (located by measurement of catalase) sedimented at 10 min. Most of the NADPH-cytochrome oxidoreductase, IDPase and acid -nitrophenyl phosphatase were still present in the supernatant. This supernatant also contained a cytochrome cytochromes -450 and -416. Zonal centrifugation indicated that catalase and cytochrome oxidase were entirely sedimentable at 6 × 10 min and that NADPH- and antimycin A-insensitive NADH-cytochrome oxidoreductases were not mitochondrial. Peroxisomes (ρ = 1.25) were separated from mitochondria (ρ = 1.20) by both rate and isopycnic-zonal centrifugation. Two zones of acid -nitrophenyl phosphatase activity were detected, one at ρ = 1.15 and the other at ρ = 1.21, but no other acid hydrolases were found. Separation, of polysomes and various microsomal enzymes (IDPase, acid -nitrophenyl phosphatase, NADH- and NADPH-cytochrome oxidoreductases) was also achieved.


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