Modeling the active sites in metalloenzymes. 3. Density functional calculations on models for [Fe]-hydrogenase: Structures and vibrational frequencies of the observed redox forms and the reaction mechanism at the diiron active center

Optimized structures for the redox species of the diiron active site in [Fe ]-hydrogenase as observed by FTLR and for species in the catalytic cycle fo r the reversible H-2 oxidation have been determined by density- functiona' calculations on the active site model, [(L)(CO)(CN)Fe(mu -PDT)(mu -CO)Fe(CO )(CN)(L ')](q) (L = H2O, CO, H-2, H- PDT = SCH2CH2CH2S, L ' = CH3S-, CH3SH; q = 0, 1-, 2-, 3-). Analyticai DFT frequencies on model complexes (mu -PDT )Fe-2(CO)(6) and [(mu -PDT)Fe-2(CO)(4)(CN)(2)](2-) are used to talibrate th e calculated CN- and CO frequencies against the measured FTIR bands in thes e model compounds. By comparing the predicted CN- and CO frequencies from D FT frequency calculations on the active site model with the observed bands of D. vulgaris [Fe]-hydrogenase under various conditions, the oxidation sta tes and structures for the diiron active site are proposed. The fully oxidi zed, EPR-silent form is an Fe(II)-Fe(II) species. Coordination of H2O to th e empty site in the enzyme's diiron active center results in an oxidized in active form (H2O)Fe(II))-Fe(II). The calculations show that reduction of th is inactive form releases the H2O to provide an open coordination site for H-2. The partially oxidized active state, which has an S = 1/2 EPR Signal, is an; Fe(I)-Fe(II) species. Fe(I)-Fe(I) species with and without bridging CO account for the fully reduced, EPR-silent state. For this fully reduced state.;the species without the bridging CO is slightly more stable than the structure with the bridging CO,The correlation coefficient between the pre dicted CN- and CO frequencies forth; proposed! model species and the measur ed CN- and CO frequencies in the enzyme is 0.964. The proposed species are so consistent with the EPR, ENDOR, and Mossbauer spectroscopies for the enz yme states.' Our results preclude the presence of Fe(III)-Fe(II) or Fe(III) -Fe(III) states among those observed by FTIR. A proposed reaction mechanism (catalytic cycle) based on the DFT calculations shows that heterolytic cle avage of H-2 can occur from (eta (2)-H-2)Fe(II)-Fe(II) via a proton transfe r to "spectator" ligands. Proton transfer to a CN- ligand is thermodynamica lly favored bur kinetically unfavorable over proton transfer to the: bridgi ng S of the PDT. Proton mi,oration from a metal hydride to a base (S, CN, o r basic protein site) results in a two-electron reduction at the metals and explains in part the active site's dimetal requirement and ligand framewor k which supports low-oxidation-state metals. The calculations also suggest that species with a protonated Fe-Fe bond could be involved if the protein could accommodate such species.