Characterization of [4Fe-4S]2+, [4Fe-4Se]2+ and hybrid (S, Se) clusters in Clostridium pasteurianum ferredoxin. A resonance Raman study

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Abstract
Resonance Raman spectra of native 2[4Fe-4S] ferredoxin from C. pasteurianum and of its Se-substituted analog 2[4Fe-4Se] are compared. The experimental conditions used in this study included low temperature and the absence of a glass sample cell, ensuring high signal-to-noise ratios. The spectra of the 2[4Fe-4S] and 2[4Fe-4Se] ferredoxins display similar numbers of bands, but the resonance Raman patterns differ largely, except for 2 bands observed at 353 cm-1 and 365 cm-1 in spectra of the native ferredoxin, which are only moderately shifted upon S .fwdarw. Se substitution and are attributed to Fe-S(cysteine) stretching modes. The activities of the latter modes, enhanced by 457.9 nm excitation, are nearly equal in both ferredoxins. These data were used to demonstrate the presence of hybrid clusters [4Fe-(4-n)S-nSe] (n = 1, 2, 3) in a ferredoxin the active sites of which were reconstituted in the presence of S and Se.

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