Electrochemical evidence on the molten globule conformation of cytochrome c.
To explore a new approach for characterizing the molten globule conformation, cyclic voltammetric studies of salt induced transitions at acidic pH of cyt c have been carried out. The use of modified electrodes has made the observation of direct electrochemistry in native cyt c possible. However, most of these electrodes do not show reversible responses at acidic pH, due to the fact that, for this system, a deprotonated electrode surface is needed. In these studies, we have used a 6-mercaptopurine and cysteine-modified gold electrodes which are effective for direct rapid electron transfer to cyt c, even in acid solutions. The change in the absorption bands of cyt c are used to monitor the conformational states and, hence, to compare the voltammetric results. Under the experimental conditions where the A state of cyt c is obtained, a reversible voltammetric signal is observed. The midpoint peak potentials are found to be very close to the formal potential of native cyt c. Results are discussed in terms of a cooperative two-state transition 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 redox properties.
Pineda
T
T
Departamento de Química Física y Termodinámica Aplicada, Facultad de Ciencias, Universidad de Córdoba, Spain.
Sevilla
J M
JM
Román
A J
AJ
Blázquez
M
M
D000818:Animals; D000838:Anions / Q000494:pharmacology; D003545:Cysteine / Q000378:metabolism; D003574:Cytochrome c Group / Q000737:chemistry*; D004563:Electrochemistry / Q000379:methods; D004566:Electrodes; D006736:Horses; D006863:Hydrogen-Ion Concentration; D015122:Mercaptopurine / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry; D010084:Oxidation-Reduction; D010472:Perchlorates / Q000494:pharmacology; D011487:Protein Conformation; D017510:Protein Folding; D017670:Sodium Compounds / Q000494:pharmacology; D013053:Spectrophotometry
D000838:Anions; D003574:Cytochrome c Group; D010472:Perchlorates; D017670:Sodium Compounds; C031068:sodium perchlorate; D015122:Mercaptopurine; D003545:Cysteine