title: [NiFe]-hydrogenase maturation: isolation of a HypC-HypD complex carrying diatomic CO and CN- ligands.
abstract: The HypC and HypD maturases are required for the biosynthesis of the Fe(CN)(2)CO cofactor in the large subunit of [NiFe]-hydrogenases. Using infrared spectroscopy we demonstrate that an anaerobically purified, Strep-tagged HypCD complex from Escherichia coli exhibits absorption bands characteristic of diatomic CO and CN(-) ligands as well as CO(2). Metal and sulphide analyses revealed that along with the [4Fe-4S](2+) cluster in HypD, the complex has two additional oxygen-labile Fe ions. We prove that HypD cysteine 41 is required for the coordination of all three ligands. These findings suggest that the HypCD complex carries minimally the Fe(CN)(2)CO cofactor.
authors.affiliation: Institute of Microbiology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany.
authors.forename: Basem
authors.initials: B
authors.lastname: Soboh
authors.forename: Sven T
authors.initials: ST
authors.lastname: Stripp
authors.forename: Enrico
authors.initials: E
authors.lastname: Muhr
authors.forename: Claudia
authors.initials: C
authors.lastname: Granich
authors.forename: Mario
authors.initials: M
authors.lastname: Braussemann
authors.forename: Martin
authors.initials: M
authors.lastname: Herzberg
authors.forename: Joachim
authors.initials: J
authors.lastname: Heberle
authors.forename: R
authors.initials: R
authors.lastname: Gary Sawers
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002248:Carbon Monoxide / Q000737:chemistry / Q000378:metabolism*; D002413:Cations, Divalent; D003067:Coenzymes / Q000737:chemistry / Q000378:metabolism; D003486:Cyanides / Q000737:chemistry / Q000378:metabolism*; 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*; D007501:Iron / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D010957:Plasmids; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013055:Spectrophotometry, Infrared
chemical_list: D000596:Amino Acids; D002413:Cations, Divalent; D003067:Coenzymes; D003486:Cyanides; D029968:Escherichia coli Proteins; C491587:HypC protein, E coli; C108875:HypD protein, Bacteria; D008024:Ligands; D011506:Proteins; D011994:Recombinant Proteins; D002248:Carbon Monoxide; D007501:Iron; C043296:nickel-iron hydrogenase; D006864:Hydrogenase