Chemical and physical characteristics of the deoxycholate-soluble and magnesium-reaggregated membrane nicotinamide adenine dinucleotide (reduced form) oxidase of Bacillus megaterium.
The inactive components of the nicotinamide adenine dinucleotide (reduced form) (NADH) oxidase present in the 0.4% deoxycholate-soluble fraction obtained from Bacillus megaterium KM membranes were reaggregated into active NADH oxidase by dilution in the presence of Mg(2+). The reaggregated oxidase was different from the original membrane with respect to sedimentation behavior in a sucrose gradient and morphological appearance. The deoxycholate-insoluble portion of the membrane had membrane-like structure whereas the reaggregated oxidase appeared to be a filamentous aggregate of small particles. The reaggregated oxidase and the deoxycholate-insoluble membrane residue were similar to the original membrane with respect to total protein and total lipid content. The inactive components of the NADH oxidase system exist in deoxycholate as two molecular species which were separable by sucrose density gradient centrifugation or gel filtration in deoxycholate-containing solutions. Both components and dilution in the presence of Mg(2+) were necessary for restoration of oxidase activity. The smaller-molecular-weight component contained all of the NADH-2,6-dichlorophenolindophenol oxidoreductase activity of the original membrane.
D001410:Bacillus megaterium / Q000032:analysis / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D001426:Bacterial Proteins / Q000032:analysis; D001647:Bile Acids and Salts; D002374:Catalase / Q000378:metabolism; D002462:Cell Membrane / Q000032:analysis / Q000201:enzymology*; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D002850:Chromatography, Gel; D006601:Hexoses / Q000032:analysis; D006885:Hydroxybutyrates / Q000032:analysis; D007215:Indophenol; D008055:Lipids / Q000032:analysis; D008274:Magnesium; D008854:Microscopy, Electron; D008970:Molecular Weight; D009243:NAD; D010088:Oxidoreductases / Q000032:analysis / Q000378:metabolism; D010758:Phosphorus / Q000032:analysis; D010772:Phosphotungstic Acid; D011523:Protoplasts; D012329:RNA, Bacterial / Q000032:analysis; D012997:Solvents; D013194:Staining and Labeling; D013395:Sucrose
D001426:Bacterial Proteins; D001647:Bile Acids and Salts; D006601:Hexoses; D006885:Hydroxybutyrates; D008055:Lipids; D012329:RNA, Bacterial; D012997:Solvents; D009243:NAD; D010772:Phosphotungstic Acid; D010758:Phosphorus; D007215:Indophenol; D013395:Sucrose; D010088:Oxidoreductases; D002374:Catalase; D008274:Magnesium