title: Subcellular fractionation by zonal centrifugation of glucose-repressed anaerobically grown Saccharomyces carlsbergensis.
abstract: 1. Homogenates were prepared from sphaeroplasts of anaerobically grown, glucoserepressed Saccharomyces carlsbergensis, and the distributions of marker enzymes investigated after zonal centrifugation on sucrose gradients containing 2mm-MgCl(2). 2. These homogenates contained no detectable cytochrome c oxidase, succinate-cytochrome c oxidoreductase, succinate-ferricyanide oxidoreductase, l(+)-lactate-cytochrome c oxidoreductase or catalase. Cytochromes a+a(3) and c were not detected. 3. Zonal centrifugation of whole homogenates indicated complex density distributions of the sedimentable portions of NADH- and NADPH-cytochrome c oxidoreductases, adenosine triphosphatases (ATPases), adenosine pyrophosphatase (ADPase), pyrophosphatase and acid p-nitrophenyl phosphatase. Several different ATPases were distinguished on the basis of their sensitivities to oligomycin and ouabain. 4. Differential centrifugation of whole homogenates at 10(5)g-min left 80-90% of the protein, dithionite-reducible cytochrome b, acid hydrolases and pyrophosphatase in a supernatant (S(1)) together with 65 and 56% of the NADH- and NADPH-cytochrome c oxidoreductases respectively, 25% of the ATPases and 71% of the adenosine monophosphatase. 5. Further analysis of supernatant S(1) revealed the presence of a class of small particles containing NADPH-cytochrome c oxidoreductases and ATPases. 6. At least four different populations of large particles were distinguished. 7. Electron microscopy indicated that one of these corresponded to ;promitochondria' as described by other workers.
mesh_terms: D000251:Adenosine Triphosphatases / Q000032:analysis; D000693:Anaerobiosis; D002458:Cell Fractionation; D002501:Centrifugation, Zonal; D003580:Cytochromes / Q000032:analysis; D005947:Glucose; D008854:Microscopy, Electron; D009243:NAD / Q000032:analysis; D009249:NADP / Q000032:analysis; D010088:Oxidoreductases / Q000032:analysis; D012440:Saccharomyces; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D003580:Cytochromes; D009243:NAD; D009249:NADP; D010088:Oxidoreductases; D000251:Adenosine Triphosphatases; D005947:Glucose