title: Development of a cell-free system reveals an oxygen-labile step in the maturation of [NiFe]-hydrogenase 2 of Escherichia coli.
abstract: By combining extracts from strains lacking genes encoding either the maturation enzymes or the large subunits of hydrogenases 1, 2 and 3 we could reconstitute in vitro under strictly anaerobic conditions 10-15% of the hydrogenase activity present in wild type Escherichia coli extracts. Purified, unprocessed Strep-tagged variants of the hydrogenase 2 large subunit, HybC, isolated from either a ΔhybD (encoding the hydrogenase 2-specific protease) mutant or a strain deficient in HypF could also be matured to active, processed enzyme using this system. These studies reveal that minimally one step early on the hydrogenase maturation pathway is oxygen-labile.
authors.affiliation: Institute for Mikrobiologie, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany.
authors.forename: Basem
authors.initials: B
authors.lastname: Soboh
authors.forename: Sara
authors.initials: S
authors.lastname: Krüger
authors.forename: Martin
authors.initials: M
authors.lastname: Kuhns
authors.forename: Constanze
authors.initials: C
authors.lastname: Pinske
authors.forename: Anne
authors.initials: A
authors.lastname: Lehmann
authors.forename: R Gary
authors.initials: RG
authors.lastname: Sawers
mesh_terms: D019878:Carboxyl and Carbamoyl Transferases / Q000235:genetics; D010450:Endopeptidases / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006864:Hydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; C491587:HypC protein, E coli; D006864:Hydrogenase; C481721:hydrogenase 2, E coli; D019878:Carboxyl and Carbamoyl Transferases; C504838:hypF protein, E coli; D010450:Endopeptidases; C119558:hydrogenase maturating endopeptidase HYBD; D010100:Oxygen