title: The specificity of proton-translocating transhydrogenase for nicotinamide nucleotides.
abstract: In its forward direction, transhydrogenase couples the reduction of NADP(+) by NADH to the outward translocation of protons across the membrane of bacteria and animal mitochondria. The enzyme has three components: dI and dIII protrude from the membrane and dII spans the membrane. Hydride transfer takes place between nucleotides bound to dI and dIII. Studies on the kinetics of a lag phase at the onset of a "cyclic reaction" catalysed by complexes of the dI and dIII components of transhydrogenase from Rhodospirillum rubrum, and on the kinetics of fluorescence changes associated with nucleotide binding, reveal two features. Firstly, the binding of NADP(+) and NADPH to dIII is extremely slow, and is probably limited by the conversion of the occluded to the open state of the complex. Secondly, dIII can also bind NAD(+) and NADH. Extrapolating to the intact enzyme this binding to the "wrong" site could lead to slip: proton translocation without change in the nucleotide redox state, which would have important consequences for bacterial and mitochondrial metabolism.
authors: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors: Lucinda
authors: L
authors: Huxley
authors: Philip G
authors: PG
authors: Quirk
authors: Nick P J
authors: NP
authors: Cotton
authors: Scott A
authors: SA
authors: White
authors: J Baz
authors: JB
authors: Jackson
mesh_terms: D019943:Amino Acid Substitution; D001665:Binding Sites; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D009250:NADP Transhydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009536:Niacinamide / Q000378:metabolism*; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012247:Rhodospirillum rubrum / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D011994:Recombinant Proteins; D009243:NAD; D009536:Niacinamide; D009249:NADP; D009250:NADP Transhydrogenases