We have designed new trithiols Temp(SH)(3) and Tefp(SH)(3) that can be synthesized conveniently in short steps and are useful for preparation of crystalline [3:1] site-differentiated [4Fe-4S] clusters suitable for X-ray structural analysis. The ethanethiolate clusters (PPh(4))(2)[Fe(4)S(4)(SEt)(TempS(3))] (4a) and (PPh(4))(2)[Fe(4)S(4)(SEt)(TefpS(3))] (4b) were prepared as precursors, and the unique iron sites were then selectively substituted. Upon reaction with H(2)S, (PPh(4))(2)[Fe(4)S(4)(SH)(TempS(3))] (6a) and (PPh(4))(2)[Fe(4)S(4)(SH)(TefpS(3))] (6b), which model the [4Fe-4S] cluster in the β subunit of (R)-2-hydroxyisocaproyl-CoA dehydratase, were synthesized. Clusters 6a and 6b were further converted to the sulfido-bridged double cubanes (PPh(4))(4)[{Fe(4)S(4)(TempS(3))}(2)(μ(2)-S)] (7a) and (PPh(4))(4)[{Fe(4)S(4)(TefpS(3))}(2)(μ(2)-S)] (7b), respectively, via intermolecular condensation with the release of H(2)S. Conversely, addition of H(2)S to 7a,b afforded the hydrosulfide clusters 6a,b. The molecular structures of the clusters reported herein were elucidated by X-ray crystallographic analysis. Their redox properties were investigated by cyclic voltammetry.

[4Fe-4S] clusters modeled after those in organisms having three cysteine thiolates and one carboxylate were synthesized by using the tridentate thiolato chelate. X-ray structural analysis reveals that the carboxylates coordinate to the unique irons in an η(1) manner rather than η(2). Redox potentials show a positive shift from that of the cluster having ethanethiolate and the tridentate thiolato chelate. These properties conform to the arrangement of the [4Fe-4S] clusters in the electron transfer systems included in Rc dark operative protochlorophyllide oxidoreductase (DPOR) and formaldehyde oxidoreductase (FOR) with Pf ferredoxin.