title: Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [(57)Fe2(adt)(CN)2(CO)4](2-).
abstract: The preparation and spectroscopic characterization of a CO-inhibited [FeFe] hydrogenase with a selectively (57)Fe-labeled binuclear subsite is described. The precursor [(57)Fe2(adt)(CN)2(CO)4](2-) was synthesized from the (57)Fe metal, S8, CO, (NEt4)CN, NH4Cl, and CH2O. (Et4N)2[(57)Fe2(adt)(CN)2(CO)4] was then used for the maturation of the [FeFe] hydrogenase HydA1 from Chlamydomonas reinhardtii, to yield the enzyme selectively labeled at the [2Fe]H subcluster. Complementary (57)Fe enrichment of the [4Fe-4S]H cluster was realized by reconstitution with (57)FeCl3 and Na2S. The Hox-CO state of [2(57)Fe]H and [4(57)Fe-4S]H HydA1 was characterized by Mössbauer, HYSCORE, ENDOR, and nuclear resonance vibrational spectroscopy.
authors.affiliation: †School of Chemical Sciences, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
authors.forename: Ryan
authors.initials: R
authors.lastname: Gilbert-Wilson
authors.affiliation: ‡Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Judith F
authors.initials: JF
authors.lastname: Siebel
authors.affiliation: ‡Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Adamska-Venkatesh
authors.affiliation: §Department of Chemistry, University of California, Davis, California 95616, United States.
authors.forename: Cindy C
authors.initials: CC
authors.lastname: Pham
authors.affiliation: ‡Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Edward
authors.initials: E
authors.lastname: Reijerse
authors.affiliation: §Department of Chemistry, University of California, Davis, California 95616, United States.|∥Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States.
authors.forename: Hongxin
authors.initials: H
authors.lastname: Wang
authors.affiliation: §Department of Chemistry, University of California, Davis, California 95616, United States.|∥Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Cramer
authors.affiliation: ‡Max-Planck-Institut für Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lubitz
authors.affiliation: †School of Chemical Sciences, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Rauchfuss
mesh_terms: D002248:Carbon Monoxide / Q000378:metabolism; D020134:Catalytic Domain; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D006864:Hydrogenase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D058085:Iron Compounds / Q000737:chemistry*; D007503:Iron Isotopes / Q000737:chemistry; D007506:Iron-Sulfur Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D015204:Spectroscopy, Mossbauer
chemical_list: D058085:Iron Compounds; D007503:Iron Isotopes; D007506:Iron-Sulfur Proteins; D002248:Carbon Monoxide; C118830:iron hydrogenase; D006864:Hydrogenase