title: Kinetics and interaction studies between cytochrome c3 and Fe-only hydrogenase from Desulfovibrio vulgaris Hildenborough.
abstract: Hydrogenases from Desulfovibrio are found to catalyze hydrogen uptake with low potential multiheme cytochromes, such as cytochrome c3, acting as acceptors. The production of Fe-only hydrogenase from Desulfovibrio vulgaris Hildenborough was improved with respect to the growth phase and media to determine the best large-scale bacteria growth conditions. The interaction and electron transfer from Fe-only hydrogenase to multiheme cytochrome has been studied in detail by both BLAcore and steady-state measurements. The electron transfer between [Fe] hydrogenase and cytochrome c3 appears to be a cooperative phenomenon (h = 1.37). This behavior could be related to the conductivity properties of multihemic cytochromes. An apparent dissociation constant was determined (2 x 10(-7) M). The importance of the cooperativity for contrasting models proposed to describe the functional role of the hydrogenase/cytochrome c3 complex is discussed. Presently, the only determined structure is from [NiFe] hydrogenase and there are no obvious similarities between [NiFe] and [Fe] hydrogenase. Furthermore, no crystallographic data are available concerning [Fe] hydrogenase. The first results on crystallization and X-ray crystallography are reported.
authors.affiliation: Laboratoire de Bioénergétique et Ingéniérie des Protéines, C.N.R.S., Institut de Biologie Structurale et Microbiologie, Marseille, France.
authors.forename: M
authors.initials: M
authors.lastname: Brugna
authors.forename: M T
authors.initials: MT
authors.lastname: Giudici-Orticoni
authors.forename: S
authors.initials: S
authors.lastname: Spinelli
authors.forename: K
authors.initials: K
authors.lastname: Brown
authors.forename: M
authors.initials: M
authors.lastname: Tegoni
authors.forename: M
authors.initials: M
authors.lastname: Bruschi
mesh_terms: D015374:Biosensing Techniques; D018360:Crystallography, X-Ray; D003574:Cytochrome c Group / Q000302:isolation & purification / Q000378:metabolism*; D016969:Desulfovibrio vulgaris / Q000378:metabolism*; D004579:Electron Transport; D006864:Hydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D011487:Protein Conformation; D017433:Protein Structure, Secondary
chemical_list: D003574:Cytochrome c Group; C017165:cytochrome c(3); D006864:Hydrogenase