ELECTROCHEMICAL EVIDENCE ON THE MOLTEN GLOBULE CONFORMATION OF CYTOCHROME-C

To explore a new approach for characterizing the molten globule confor mation, cyclic voltammetric studies of salt induced transitions at aci dic pH of cyt c have been carried out. The use of modified electrodes has made the observation of direct electrochemistry in native cyt c po ssible. However, most of these electrodes do not show reversible respo nses at acidic pH, due to the fact that, for this system, a deprotonat ed electrode surface is needed. Tn these studies, we have used a 6-mer captopurine and cysteine-modified gold electrodes which are effective for direct rapid electron transfer to cyt c, even in acid solutions. T he change in the absorption bands of cyt c are used to monitor the con formational states and, hence, to compare the voltammetric results. Un der the experimental conditions where the A state of cyt c is obtained , a reversible voltammetric signal is observed. The midpoint peak pote ntials are found to be very close to the formal potential of native cy t c. Results are discussed in terms of a cooperative two-state transit ion between the acid unfolded and the globular acidic states of cyt c. This finding establishes, for the first time, the similarity of both the native and the molten globule-like conformations in terms of its r edox properties. (C) 1997 Elsevier Science B.V.