title: Properties of the respiratory NAD(P)H dehydrogenase isolated from the cyanobacterium Synechocystis PCC6803.
abstract: Activity staining with NADPH-nitroblue tetrazolium after native-PAGE of membrane proteins of Synechocystis PCC6803, solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), revealed four NAD(P)H dehydrogenase (NDH) activities; an NDH complex of the respiratory chain, a ferredoxin NADP+ reductase (FNR), a drgA product which oxidized both NADH and NADPH, and an uncharacterized NADH-specific enzyme. The NDH complex was purified with anion exchange and gel filtration chromatographies. The purified complex had a molecular mass of 376 kDa and was composed of 9 subunits. Western analysis showed that the complex contained the NDH-H subunit, but not NDH-A or B. The enzyme reduced ferricyanide much faster than plastoquinone and used NADPH as its preferred electron donor rather than NADH. The enzymatic activity was inhibited by diphenyleneiodonium chloride and salicylhydroxamic acid, but not by rotenone, p-chloromercuribenzoate, N-ethylmaleimide, flavon, dicumarol, or antimycin A. These results suggest that the purified complex is a hydrophilic subcomplex which contains an NADPH binding site and flavin, and is dissociated from a hydrophobic subcomplex, which contains quinone binding site.
authors.affiliation: Research Institute for Food Science, Kyoto University, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Matsuo
authors.forename: T
authors.initials: T
authors.lastname: Endo
authors.forename: K
authors.initials: K
authors.lastname: Asada
mesh_terms: D000595:Amino Acid Sequence; D000458:Cyanobacteria / Q000201:enzymology*; D004579:Electron Transport; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010101:Oxygen Consumption; D010446:Peptide Fragments / Q000276:immunology; D011487:Protein Conformation
chemical_list: D010446:Peptide Fragments; D009243:NAD; D009249:NADP; D016660:NAD(P)H Dehydrogenase (Quinone)