purple acidophilic di-ferric DNA ligase from Ferroplasma

• Published in: Proceedings of the National Academy of Sciences - PNAS 2008-07, Vol.105 (26), p.8878-8883
• Main Authors: Ferrer, Manuel, Golyshina, Olga V, Beloqui, Ana, Böttger, Lars H, Andreu, José M, Polaina, Julio, De Lacey, Antonio L, Trautwein, Alfred X, Timmis, Kenneth N, Golyshin, Peter N
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Archaea; Binding sites; Biochemistry; Biological Sciences; Colors; Deoxyribonucleic acid; DNA; DNA - metabolism; DNA Ligase ATP; DNA Ligases - chemistry; DNA Ligases - metabolism; Enzymes; Ferroplasma; Genomes; Hydrogen-Ion Concentration; Iron; Iron - metabolism; Ligation; Metal ions; Molecular Sequence Data; Mutation; Protein Conformation; Protein folding; Recombinant Proteins - metabolism; Spectroscopy, Mossbauer; Substrate Specificity; Thermoplasmales; Thermoplasmales - enzymology
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0800071105

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 Fe³⁺-tyrosinate centers and lacks any requirement for either Mg²⁺ or K⁺ for activity. DNA ligases from closest phylogenetic and ecophysiological relatives have normal pH optima (6.0-7.5), lack iron, and require Mg²⁺/K⁺ for activity. Ferric iron retention is pH-dependent, with release resulting in partial protein unfolding and loss of activity. Reduction of the Fe³⁺ to Fe²⁺ 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.