Structural redox control in a 7Fe ferredoxin isolated from Desulfovibrio alaskensis

Abstract

The redox behaviour of a ferredoxin (Fd) from Desulfovibrio alaskensis was characterized by electrochemistry. The protein was isolated and purified, and showed to be a tetramer containing one [3Fe–4S] and one [4Fe–4S] centre. This ferredoxin has high homology with FdI from Desulfovibrio vulgaris Miyazaki and Hildenborough and FdIII from Desulfovibrio africanus. From differential pulse voltammetry the following signals were identified: [3Fe-4S] + 1/0 (E 0′ = − 158 ± 5 mV); [4Fe–4S] + 2/+1 (E 0′ = − 474 ± 5 mV) and [3Fe–4S] 0/− 2 (E 0′ = − 660 ± 5 mV). The effect of pH on these signals showed that the reduced [3Fe–4S] 0 cluster has a p Kʹ red ′ = 5.1 ± 0.1, the [4Fe–4S] + 2/+1 centre is pH independent, and the [3Fe–4S] 0/−2 reduction is accompanied by the binding of two protons. The ability of the [3Fe–4S] 0 cluster to be converted into a new [4Fe–4S] cluster was proven. The redox potential of the original [4Fe–4S] centre showed to be dependent on the formation of the new [4Fe-4S] centre, which results in a positive shift ( ca. 70 mV) of the redox potential of the original centre. Being most [Fe–S] proteins involved in electron transport processes, the electrochemical characterization of their clusters is essential to understand their biological function. Complementary EPR studies were performed.