title: An all sulfur analogue of the smallest subunit of F420-non-reducing hydrogenase from Methanococcus voltae--metal binding and structure.
abstract: The 25 amino acid long subunit VhuU of the F420-non-reducing hydrogenase from Methanococcus voltae contains selenocysteine within the consensus sequence of known [NiFe] hydrogenases DP(C or U)CxxCxxH (U = selenocysteine). The sulfur-analogue VhuUc was chemically synthesized, purified and its metal binding capability, the catalytic properties, and structural features were investigated. The polypeptide was able to bind nickel, but did not catalyse the heterolytic activation of H2. 2D-NMR spectroscopy revealed an alpha-helical secondary structure for the 15 N-terminal amino acids in 50% TFE. Nickel only binds to the C-terminus, which contains the conserved amino acid motif. Structures derived from the NMR data are compatible with the participation of both sulfur atoms from the conserved cysteine residues in a metal ion binding. Structures obtained from the data sets for Ni.VhuUc as well as Zn.VhuUc showed no further ligands. The informational value for Ni.VhuUc was low due to paramagnetism.
authors.affiliation: Department of Biology, University of Marburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Pfeiffer
authors.forename: A
authors.initials: A
authors.lastname: Klein
authors.forename: P
authors.initials: P
authors.lastname: Steinert
authors.forename: D
authors.initials: D
authors.lastname: Schomburg
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002384:Catalysis; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008670:Metals / Q000378:metabolism*; D017017:Methanococcus / Q000201:enzymology*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009532:Nickel / Q000378:metabolism; D017433:Protein Structure, Secondary; D017279:Selenocysteine / Q000032:analysis; D016415:Sequence Alignment; D013455:Sulfur
chemical_list: D008670:Metals; D017279:Selenocysteine; D013455:Sulfur; D009532:Nickel; D006859:Hydrogen; C058560:nickel-iron-selenium hydrogenase; D006864:Hydrogenase