Specific ion effects on the electrochemical properties of cytochromec

The range of salts used as supporting electrolytes in electrochemical studies of redox proteins and enzymes varies widely, with the choice of an electrolyte relying on the assumption that the electrolyte used does not affect the electrochemical properties of the proteins and enzymes under investigation. Examination of the electrochemical properties of the redox protein cytochrome c (cyt c) at a 4,4′-bipyridyl modified gold electrode demonstrates that both the redox potential (Eo′) and the faradaic current are influenced by the nature of the electrolyte used, in a manner explained primarily by Hofmeister effects. The faradaic peak currents display an atypical trend on switching from kosmotropic to chaotropic anions, with a maximum current observed in the presence of Cl−. For a series of cations, the peak current increased in the sequence: Li+ (0.34 μA) < guanidinium+ (0.36 μA) < Na+ (0.37 μA) < K+ (0.38 μA) < Cs+ (0.40 μA) and for anions it decreased in the sequence: Cl− (0.37 μA) > Br− (0.35 μA) > ClO4− (0.35 μA) > SCN− (0.31 μA) > F− (0.30 μA). Eo′ decreased by a total of 24 mV across the series F− > Cl− > Br− > ClO4− > SCN− whereas no specific ion effect on Eo′ was observed for cations. Factorisation of Eo′ into its enthalpic and entropic components showed that while no specific trends were observed, large changes in ΔHo′ and ΔSo′ occurred with individual ions. The effect of anions on the faradaic peak current can be qualitatively explained by considering Collins' empirical rule of ‘matching water affinities’. The effect of cations cannot be explained by this rule. However, both anion and cation effects can be understood by taking into account the cooperative action of electrostatic and ion dispersion forces. The results demonstrate that the choice of a supporting electrolyte in electrochemical investigations of redox proteins is important and emphasize that care needs to be taken in the determination and comparison of Eo′, ΔHo′ and ΔSo′ in different solutions.