title: A purple acidophilic di-ferric DNA ligase from Ferroplasma.
abstract: We describe here an extraordinary purple-colored DNA ligase, LigFa, from the acidophilic ferrous iron-oxidizing archaeon Ferroplasma acidiphilum, a di-ferric enzyme with an extremely low pH activity optimum. Unlike any other DNA ligase studied to date, LigFa contains two Fe(3+)-tyrosinate centers and lacks any requirement for either Mg(2+) or K(+) for activity. DNA ligases from closest phylogenetic and ecophysiological relatives have normal pH optima (6.0-7.5), lack iron, and require Mg(2+)/K(+) for activity. Ferric iron retention is pH-dependent, with release resulting in partial protein unfolding and loss of activity. Reduction of the Fe(3+) to Fe(2+) results in an 80% decrease in DNA substrate binding and an increase in the pH activity optimum to 5.0. DNA binding induces significant conformational change around the iron site(s), suggesting that the ferric irons of LigFa act both as structure organizing and stabilizing elements and as Lewis acids facilitating DNA binding at low pH.
authors.affiliation: Consejo Superior de Investigaciones Científicas, Institute of Catalysis, 28049 Madrid, Spain. mferrer@icp.csic.es
authors.forename: Manuel
authors.initials: M
authors.lastname: Ferrer
authors.forename: Olga V
authors.initials: OV
authors.lastname: Golyshina
authors.forename: Ana
authors.initials: A
authors.lastname: Beloqui
authors.forename: Lars H
authors.initials: LH
authors.lastname: Böttger
authors.forename: José M
authors.initials: JM
authors.lastname: Andreu
authors.forename: Julio
authors.initials: J
authors.lastname: Polaina
authors.forename: Antonio L
authors.initials: AL
authors.lastname: De Lacey
authors.forename: Alfred X
authors.initials: AX
authors.lastname: Trautwein
authors.forename: Kenneth N
authors.initials: KN
authors.lastname: Timmis
authors.forename: Peter N
authors.initials: PN
authors.lastname: Golyshin
mesh_terms: D000595:Amino Acid Sequence; D004247:DNA / Q000378:metabolism; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D015204:Spectroscopy, Mossbauer; D013379:Substrate Specificity; D019603:Thermoplasmales / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D004247:DNA; D007501:Iron; D011088:DNA Ligases; D000072481:DNA Ligase ATP