title: Crystal structure of HydF scaffold protein provides insights into [FeFe]-hydrogenase maturation.
abstract: [FeFe]-hydrogenases catalyze the reversible production of H2 in some bacteria and unicellular eukaryotes. These enzymes require ancillary proteins to assemble the unique active site H-cluster, a complex structure composed of a 2Fe center bridged to a [4Fe-4S] cubane. The first crystal structure of a key factor in the maturation process, HydF, has been determined at 3 Å resolution. The protein monomer present in the asymmetric unit of the crystal comprises three domains: a GTP-binding domain, a dimerization domain, and a metal cluster-binding domain, all characterized by similar folding motifs. Two monomers dimerize, giving rise to a stable dimer, held together mainly by the formation of a continuous β-sheet comprising eight β-strands from two monomers. Moreover, in the structure presented, two dimers aggregate to form a supramolecular organization that represents an inactivated form of the HydF maturase. The crystal structure of the latter furnishes several clues about the events necessary for cluster generation/transfer and provides an excellent model to begin elucidating the structure/function of HydF in [FeFe]-hydrogenase maturation.
authors: Department of Biological Chemistry, University of Padua, 35131 Padua, Italy.
authors: Laura
authors: L
authors: Cendron
authors: Department of Biology, University of Padua, 35131 Padua, Italy.
authors: Paola
authors: P
authors: Berto
authors: Department of Biology, University of Padua, 35131 Padua, Italy; Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, Colorado 80401.
authors: Sarah
authors: S
authors: D'Adamo
authors: Department of Biology, University of Padua, 35131 Padua, Italy.
authors: Francesca
authors: F
authors: Vallese
authors: Department of Biotechnologies, University of Verona, 37134 Verona, Italy.
authors: Chiara
authors: C
authors: Govoni
authors: Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, Colorado 80401.
authors: Matthew C
authors: MC
authors: Posewitz
authors: Department of Biology, University of Padua, 35131 Padua, Italy.
authors: Giorgio M
authors: GM
authors: Giacometti
authors: Department of Biology, University of Padua, 35131 Padua, Italy. Electronic address: paola.costantini@unipd.it.
authors: Paola
authors: P
authors: Costantini
authors: Department of Biological Chemistry, University of Padua, 35131 Padua, Italy. Electronic address: giuseppe.zanotti@unipd.it.
authors: Giuseppe
authors: G
authors: Zanotti
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D002417:Cattle; D018360:Crystallography, X-Ray / Q000379:methods; D019281:Dimerization; D020558:GTP Phosphohydrolases / Q000737:chemistry* / Q000235:genetics; D006160:Guanosine Triphosphate / Q000737:chemistry; D006864:Hydrogenase / Q000737:chemistry*; D007501:Iron / Q000737:chemistry*; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D046949:Thermotoga neapolitana / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D006160:Guanosine Triphosphate; D007501:Iron; D006864:Hydrogenase; D020558:GTP Phosphohydrolases