title: A model system for [NiFe] hydrogenase maturation studies: Purification of an active site-containing hydrogenase large subunit without small subunit.
abstract: The large subunit HoxC of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha was purified without its small subunit. Two forms of HoxC were identified. Both forms contained iron but only substoichiometric amounts of nickel. One form was a homodimer of HoxC whereas the second also contained the Ni-Fe site maturation proteins HypC and HypB. Despite the presence of the Ni-Fe active site in some of the proteins, both forms, which lack the Fe-S clusters normally present in hydrogenases, cannot activate hydrogen. The incomplete insertion of nickel into the Ni-Fe site provides direct evidence that Fe precedes Ni in the course of metal center assembly.
authors.affiliation: Institut für Biologie/Mikrobiologie, Humboldt-Universität zu Berlin, Chausseestr. 117, D-10115 Berlin, Germany.
authors.forename: Gordon
authors.initials: G
authors.lastname: Winter
authors.forename: Thorsten
authors.initials: T
authors.lastname: Buhrke
authors.forename: Oliver
authors.initials: O
authors.lastname: Lenz
authors.forename: Anne Katherine
authors.initials: AK
authors.lastname: Jones
authors.forename: Michael
authors.initials: M
authors.lastname: Forgber
authors.forename: Bärbel
authors.initials: B
authors.lastname: Friedrich
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D001665:Binding Sites; D020118:Cupriavidus necator / Q000201:enzymology*; D019281:Dimerization; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000737:chemistry / Q000302:isolation & purification*; D007501:Iron / Q000737:chemistry; D009532:Nickel / Q000737:chemistry; D021122:Protein Subunits / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D021122:Protein Subunits; D009532:Nickel; D006859:Hydrogen; D007501:Iron; C043296:nickel-iron hydrogenase; D006864:Hydrogenase