title: Cytochrome c553 from Desulfovibrio vulgaris (Hildenborough). Electrochemical properties and electron transfer with hydrogenase.
abstract: An electrochemical study of the periplasmic cytochrome c553 of Desulfovibrio vulgaris (Hildenborough) is presented. The dependence of the midpoint potential on temperature and pH was studied with cyclic voltammetry. The voltammograms obtained were reversible and revealed that this cytochrome showed fast electron transfer on a bare glassy carbon electrode. The midpoint potential at pH 7.0 and 25 degrees C was found to be 62 mV versus the normal hydrogen electrode. It was observed that the temperature dependence was discontinuous with a transition temperature at 32 degrees C. The standard reaction entropy at the growth temperature of the organism (37 degrees C) was calculated to be delta S degree ' = -234 J mol-1 K-1. The pH dependence of the midpoint potential could be described with one pK of the oxidized form with a value of 10.6. The second-order rate constant for electron transfer between cytochrome c553 and the Fe-hydrogenase from D. vulgaris (H) was also determined with cyclic voltammetry. The equivalent rate constant for cytochrome c3 and hydrogenase was measured for comparison. The second-order rate constants are 2 x 10(7) M-1 s-1 for cytochrome c553 and 2 x 10(8) M-1 s-1 for cytochrome c3. The kinetic parameters of the hydrogenase for both cytochromes were determined using the spectrophotometric hydrogen consumption assay. With cytochrome c553 this resulted in a Km of 46 microM and a maximum turnover number of 7.1 x 10(2) s-1 in the H2 consumption assay. The values with cytochrome c3 were 17 microM and 6.4 x 10(2) s-1, respectively. The importance of the different kinetic parameters for contrasting models proposed to describe the function of the Fe-hydrogenase are discussed.
authors.affiliation: Department of Biochemistry, Wageningen Agricultural University, The Netherlands.
authors.forename: M F
authors.initials: MF
authors.lastname: Verhagen
authors.forename: R B
authors.initials: RB
authors.lastname: Wolbert
authors.forename: W R
authors.initials: WR
authors.lastname: Hagen
mesh_terms: D003574:Cytochrome c Group / Q000737:chemistry* / Q000378:metabolism; D016969:Desulfovibrio vulgaris / Q000201:enzymology*; D004563:Electrochemistry; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D006864:Hydrogenase / Q000378:metabolism*; D007700:Kinetics; D009572:Nitrite Reductases / Q000737:chemistry* / Q000378:metabolism; D009994:Osmolar Concentration; D013696:Temperature; D013816:Thermodynamics
chemical_list: D003574:Cytochrome c Group; C083951:cytochrome c553; C017165:cytochrome c(3); C022302:cytochrome C-552; D006864:Hydrogenase; D009572:Nitrite Reductases