The formate dehydrogenase-cytochrome c553 complex from Desulfovibrio vulgaris Hildenborough.

The electron transfer between formate dehydrogenase and cytochrome c553 from the anaerobic bacteria Desulfovibrio vulgaris Hildenborough has been investigated. Parameters of the electron transfer kinetics are reported. The ionic strength dependence of the complex formation has been evidenced. Two mutants of cytochrome c553 have been obtained using site-directed mutagenesis with the substitutions K62E and K62E,K63E. According to one-dimensional and two-dimensional NMR analysis, the two variants were found to have the same folding pattern as that of the wild-type cytochrome. The replacements of the lysine residues by acidic groups have important effects on the affinity between the two oxidoreduction partners. K62 and K63 are essential for recognition between the formate dehydrogenase and the cytochrome c553. Previous structural studies of cytochrome c553 have demonstrated the involvement of the polypeptide chain in the modulation of the particular low oxidoreduction potential of this cytochrome. The present study provides evidence that, during the evolution of cytochromes from the anaerobic metabolism to aerobic respiration and photosynthesis, the electrostatic distribution at the recognised encounter surface around the heme is highly conserved in all cytochromes.