Role of the tetrahemic subunit in Desulfovibrio vulgaris hildenborough formate dehydrogenase.
In the anaerobic sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough (DvH), the genome sequencing revealed the presence of three operons encoding formate dehydrogenases. fdh1 encodes an alphabetagamma trimeric enzyme containing 11 heme binding sites; fdh2 corresponds to an alphabetagamma trimeric enzyme with a tetrahemic subunit; fdh3 encodes an alphabeta dimeric enzyme. In the present work, spectroscopic measurements demonstrated that the reduction of cytochrome c(553) was obtained in the presence of the trimeric FDH2 and not with the dimeric FDH3, suggesting that the tetrahemic subunit (FDH2C) is essential for the interaction with this physiological electron transfer partner. To further study the role of the tetrahemic subunit, the fdh2C gene was cloned and expressed in Desulfovibrio desulfuricans G201. The recombinant FDH2C was purified and characterized by optical and NMR spectroscopies. The heme redox potentials measured by electrochemistry were found to be identical in the whole enzyme and in the recombinant subunit, indicating a correct folding of the recombinant protein. The mapping of the interacting site by 2D heteronuclear NMR demonstrated a similar interaction of cytochrome c(553) with the native enzyme and the recombinant subunit. The presence of hemes c in the gamma subunit of formate dehydrogenases is specific of these anaerobic sulfate-reducing bacteria and replaces heme b subunit generally found in the enzymes involved in anaerobic metabolisms.
ElAntak
Latifa
L
Unité de Bioénergétique et Ingénierie des Protéines, IBSM-CNRS, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Dolla
Alain
A
Durand
Marie-Claire
MC
Bianco
Pierre
P
Guerlesquin
Françoise
F
D000595:Amino Acid Sequence; D001665:Binding Sites; D003574:Cytochrome c Group / Q000737:chemistry; D016969:Desulfovibrio vulgaris / Q000201:enzymology*; D005560:Formate Dehydrogenases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006420:Hemeproteins / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010084:Oxidation-Reduction; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment
D003574:Cytochrome c Group; D006420:Hemeproteins; D021122:Protein Subunits; D011994:Recombinant Proteins; C083951:cytochrome c553; D005560:Formate Dehydrogenases