Hydrogenases are enzymes which catalyze the reversible cleavage of molecular hydrogen into protons and electrons. In [FeFe] hydrogenases the active center is a 6Fe6S cluster, referred to as the "H-cluster." It consists of the redox-active binuclear subcluster ([2Fe]H) coordinated by CN- and CO ligands and the cubane-like [4Fe-4S] H subcluster which is connected to the protein via Cys ligands. One of these Cys ligands bridges to the [2Fe]H subcluster. The CO-inhibited form of [FeFe] hydrogenase isolated from Desulfovibrio desulfuricans was studied using advanced EPR methods. In the Hox-CO state the open coordination site at the [2Fe]H subcluster is blocked by extrinsic CO, giving rise to an EPR-active S = 1/2 species. The CO inhibited state was prepared with 13CO and illuminated under white light at 273 K. In this case scrambling of the CO ligands occurs. Three 13C hyperfine couplings of 17.1, 7.4, and 3.8 MHz (isotropic part) were observed and assigned to 13CO at the extrinsic, the bridging, and the terminal CO-ligand positions of the distal iron, respectively. No 13CO exchange of the CO ligand to the proximal iron was observed. The hyperfine interactions detected indicate a rather large distribution of the spin density over the terminal and bridging CO ligands attached to the distal iron. Furthermore, 14N nuclear spin interactions were measured. On the basis of the observed 14N hyperfine couplings, which result from the CN- ligands of the [2Fe]H subcluster, it has been concluded that there is very little unpaired spin density on the cyanides of the binuclear subcluster.
All Science Journal Classification (ASJC) codes
- Inorganic Chemistry