Mapping the cytochrome c553 interacting site using 1H and 15N NMR.
Cytochrome c553 is the electron transfer partner of formate dehydrogenase and of [Fel-hydrogenase, two metalloenzymes essential in the metabolism of sulfate reducing bacteria. These two enzymes contain a 'ferredoxin-like' domain which presents 30% identity with Desulfovibrio desulfuricans Norway ferredoxin 1. This was chosen as a model for the 'ferredoxin-like' domain involved in the electron transfer reaction with cytochrome c553. ID NMR titration of complex formation gave us the stoichiometry (1:1) and the dissociation constant of the complex (Kd approximately 3x10(-6) M). 2D heteronuclear NMR experiments were performed to analyze the 1H and 15N chemical shift variations that are induced by the protein-protein recognition. This is the first mapping of the interaction site on a c-type cytochrome, using heteronuclear NMR.
Morelli
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Unité de Bioénergétique et Ingénierie des Protéines, IBSM-CNRS, Marseille, France.
Guerlesquin
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D001426:Bacterial Proteins / Q000737:chemistry; D001665:Binding Sites; D003574:Cytochrome c Group / Q000737:chemistry*; D003901:Desulfovibrio / Q000201:enzymology*; D005288:Ferredoxins / Q000737:chemistry; D005560:Formate Dehydrogenases / Q000737:chemistry; D006864:Hydrogenase / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular
D001426:Bacterial Proteins; D003574:Cytochrome c Group; D005288:Ferredoxins; C083951:cytochrome c553; D006864:Hydrogenase; D005560:Formate Dehydrogenases; C020644:Formate dehydrogenase (cytochrome)