title: Sulphur shuttling across a chaperone during molybdenum cofactor maturation.
abstract: Formate dehydrogenases (FDHs) are of interest as they are natural catalysts that sequester atmospheric CO2, generating reduced carbon compounds with possible uses as fuel. FDHs activity in Escherichia coli strictly requires the sulphurtransferase EcFdhD, which likely transfers sulphur from IscS to the molybdenum cofactor (Mo-bisPGD) of FDHs. Here we show that EcFdhD binds Mo-bisPGD in vivo and has submicromolar affinity for GDP-used as a surrogate of the molybdenum cofactor's nucleotide moieties. The crystal structure of EcFdhD in complex with GDP shows two symmetrical binding sites located on the same face of the dimer. These binding sites are connected via a tunnel-like cavity to the opposite face of the dimer where two dynamic loops, each harbouring two functionally important cysteine residues, are present. On the basis of structure-guided mutagenesis, we propose a model for the sulphuration mechanism of Mo-bisPGD where the sulphur atom shuttles across the chaperone dimer.
authors: 1] Laboratoire de Bioénergétique Cellulaire, CEA, DSV, IBEB, 13108, Saint-Paul-lez-Durance, France [2] CNRS, UMR 7265 Biologie Végétale et Microbiologie Environnementales, 13108 Saint-Paul-lez-Durance, France [3] Aix Marseille Université, CEA, CNRS, Biologie Végétale et Microbiologie Environnementales UMR 7265, 13108 Saint-Paul-lez-Durance, France.
authors: Pascal
authors: P
authors: Arnoux
authors: Department of Plant Biology, Technical University, Braunschweig D-38106, Germany.
authors: Christian
authors: C
authors: Ruppelt
authors: Aix Marseille Université, CNRS, IMM, LCB UMR 7283, 13402 Marseille, France.
authors: Flore
authors: F
authors: Oudouhou
authors: 1] Laboratoire de Bioénergétique Cellulaire, CEA, DSV, IBEB, 13108, Saint-Paul-lez-Durance, France [2] CNRS, UMR 7265 Biologie Végétale et Microbiologie Environnementales, 13108 Saint-Paul-lez-Durance, France [3] Aix Marseille Université, CEA, CNRS, Biologie Végétale et Microbiologie Environnementales UMR 7265, 13108 Saint-Paul-lez-Durance, France.
authors: Jérôme
authors: J
authors: Lavergne
authors: 1] Laboratoire de Bioénergétique Cellulaire, CEA, DSV, IBEB, 13108, Saint-Paul-lez-Durance, France [2] CNRS, UMR 7265 Biologie Végétale et Microbiologie Environnementales, 13108 Saint-Paul-lez-Durance, France [3] Aix Marseille Université, CEA, CNRS, Biologie Végétale et Microbiologie Environnementales UMR 7265, 13108 Saint-Paul-lez-Durance, France.
authors: Marina I
authors: MI
authors: Siponen
authors: Aix Marseille Université, CNRS, IMM, LCB UMR 7283, 13402 Marseille, France.
authors: René
authors: R
authors: Toci
authors: Department of Plant Biology, Technical University, Braunschweig D-38106, Germany.
authors: Ralf R
authors: RR
authors: Mendel
authors: Department of Plant Biology, Technical University, Braunschweig D-38106, Germany.
authors: Florian
authors: F
authors: Bittner
authors: 1] Laboratoire de Bioénergétique Cellulaire, CEA, DSV, IBEB, 13108, Saint-Paul-lez-Durance, France [2] CNRS, UMR 7265 Biologie Végétale et Microbiologie Environnementales, 13108 Saint-Paul-lez-Durance, France [3] Aix Marseille Université, CEA, CNRS, Biologie Végétale et Microbiologie Environnementales UMR 7265, 13108 Saint-Paul-lez-Durance, France.
authors: David
authors: D
authors: Pignol
authors: Aix Marseille Université, CNRS, IMM, LCB UMR 7283, 13402 Marseille, France.
authors: Axel
authors: A
authors: Magalon
authors: Aix Marseille Université, CNRS, IMM, LCB UMR 7283, 13402 Marseille, France.
authors: Anne
authors: A
authors: Walburger
mesh_terms: D001665:Binding Sites; D055162:Biocatalysis; D057486:Carbon Cycle; D002245:Carbon Dioxide / Q000378:metabolism; D013437:Carbon-Sulfur Lyases / Q000378:metabolism; D003001:Cloning, Molecular; D003067:Coenzymes / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005560:Formate Dehydrogenases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005561:Formates / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D006153:Guanosine Diphosphate / Q000737:chemistry* / Q000378:metabolism; D006864:Hydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008982:Molybdenum / Q000737:chemistry* / Q000378:metabolism; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010084:Oxidation-Reduction; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013455:Sulfur / Q000737:chemistry / Q000378:metabolism
chemical_list: D003067:Coenzymes; D005561:Formates; D018832:Molecular Chaperones; D009097:Multienzyme Complexes; C030544:formic acid; D002245:Carbon Dioxide; D006153:Guanosine Diphosphate; D013455:Sulfur; D008982:Molybdenum; D006864:Hydrogenase; D005560:Formate Dehydrogenases; C054017:formate hydrogenlyase; D013437:Carbon-Sulfur Lyases; C080546:cysteine desulfurase