title: Structure of hydrogenase maturation protein HypF with reaction intermediates shows two active sites.
abstract: [NiFe]-hydrogenases are multimeric proteins. The large subunit contains the NiFe(CN)(2)CO bimetallic active center and the small subunit contains Fe-S clusters. Biosynthesis and assembly of the NiFe(CN)(2)CO active center requires six Hyp accessory proteins. The synthesis of the CN(-) ligands is catalyzed by the combined actions of HypF and HypE using carbamoylphosphate as a substrate. We report the structure of Escherichia coli HypF(92-750) lacking the N-terminal acylphosphatase domain. HypF(92-750) comprises the novel Zn-finger domain, the nucleotide-binding YrdC-like domain, and the Kae1-like universal domain, also binding a nucleotide and a Zn(2+) ion. The two nucleotide-binding sites are sequestered in an internal cavity, facing each other and separated by ∼14 Å. The YrdC-like domain converts carbamoyl moiety to a carbamoyl adenylate intermediate, which is channeled to the Kae1-like domain. Mutations within either nucleotide-binding site compromise hydrogenase maturation but do not affect the carbamoylphosphate phosphatase activity.
authors.affiliation: Department of Biochemistry, McGill University, Montreal, QC H3G 1Y6, Canada.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Petkun
authors.forename: Rong
authors.initials: R
authors.lastname: Shi
authors.forename: Yunge
authors.initials: Y
authors.lastname: Li
authors.forename: Abdalin
authors.initials: A
authors.lastname: Asinas
authors.forename: Christine
authors.initials: C
authors.lastname: Munger
authors.forename: Linhua
authors.initials: L
authors.lastname: Zhang
authors.forename: Mandy
authors.initials: M
authors.lastname: Waclawek
authors.forename: Basem
authors.initials: B
authors.lastname: Soboh
authors.forename: R Gary
authors.initials: RG
authors.lastname: Sawers
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Cygler
mesh_terms: D017766:Acid Anhydride Hydrolases / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002221:Carbamyl Phosphate / Q000378:metabolism; D019878:Carboxyl and Carbamoyl Transferases / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D020134:Catalytic Domain; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D006864:Hydrogenase / Q000737:chemistry; D008024:Ligands; D000097024:Acylphosphatase
chemical_list: D017766:Acid Anhydride Hydrolases; D002221:Carbamyl Phosphate; D019878:Carboxyl and Carbamoyl Transferases; D029968:Escherichia coli Proteins; D006864:Hydrogenase; D008024:Ligands; D000097024:Acylphosphatase; C043296:nickel-iron hydrogenase; C504838:hypF protein, E coli