Spectroscopic and Structural Characterization of Reduced Desulfovibrio vulgaris Hildenborough W-FdhAB Reveals Stable Metal Coordination during Catalysis

Metal-dependent formate dehydrogenases are important enzymes due to their activity of CO2 reduction to formate. The tungsten-containing FdhAB formate dehydrogenase from Desulfovibrio vulgaris Hildenborough is a good example displaying high activity, simple composition, and a notable structural and catalytic robustness. Here, we report the first spectroscopic redox characterization of FdhAB metal centers by EPR. Titration with dithionite or formate leads to reduction of three [4Fe-4S](1+) clusters, and full reduction requires Ti(III)-citrate. The redox potentials of the four [4Fe-4S](1+) centers range between -250 and -530 mV. Two distinct W-V signals were detected, W-D(V) and W-F(V), which differ in only the g(2)-value. This difference can be explained by small variations in the twist angle of the two pyranopterins, as determined through DFT calculations of model compounds. The redox potential of W-VI/V was determined to be -370 mV when reduced by dithionite and -340 mV when reduced by formate. The crystal structure of dithionite-reduced FdhAB was determined at high resolution (1.5 & Aring;), revealing the same structural alterations as reported for the formate-reduced structure. These results corroborate a stable six-ligand W coordination in the catalytic intermediate W-V state of FdhAB.

Automated summary

The study investigated the structure and catalytic mechanism of metal-dependent formate dehydrogenases, revealing their high activity and structural stability. Spectroscopic and crystallographic analysis determined the redox characteristics and structural changes of the metal centers.