Conformationally gated electron transfer in iso-1-cytochrome c: engineering the rate of a conformational switch

Abstract

An engineered form of iso-1-cytochrome c with lysine 73 mutated to histidine is shown to increase by nearly 500-fold the rate of a conformational gate that modulates the rate of electron transfer into this protein. This result demonstrates the potential of protein engineering to provide electron transfer gates with tailored properties. The pH dependence of the rate of the conformational electron transfer gate correlates well with the pH dependence of the conformational change from a His 73-ligated heme to a Met 80-ligated heme, determined independently by pH jump methods, allowing unambiguous assignment of the conformational electron transfer gating step. The rate of the electron transfer gate is also modulated by a cis to trans proline isomerization, indicating that both amino acid sequence and the nature of the heme ligand provide avenues for rational design of electron transfer gates which open at different rates.