Serum cytochrome c detection using a cytochrome c oxidase biosensor.
Within the last ten years it has emerged that the release of cytochrome c plays a critical role in the important process of programmed cell death. It has also been shown that this protein is released into the circulating blood following MIs (myocardial infarctions). Methods for the detection of this protein have therefore become important. The enzyme cytochrome c oxidase is specific for cytochrome c. Bovine cytochrome c oxidase was successfully immobilized in a didodecyldimethylammonium bromide vesicular system on to gold electrodes and its interaction with cytochrome c was studied. Square-wave-voltammetric analysis of the biosensor showed two redox couples with midpoint potential, E(0)', values of +182 and +414 mV compared with Ag/AgCl. The redox couple with E(0)' of +414 mV showed a cathodic sensitivity to the presence of cytochrome c in both buffer solution and human serum. Responses of the cytochrome c oxidase biosensor to oxidized cytochrome c followed hyperbolic electrochemical Michaelis-Menten kinetics with a K(m) of 1.57 microM and maximum current (I(max)) of 1.38 x 10(-6) muA. The detection limit of the biosensor in human serum was 0.2 microM, which is well below the lowest physiological concentration of 0.8 muM previously reported for MIs [Alleyne, Joseph and Sampson (2001) Appl. Biochem. Biotechnol. 90, 97-105]. These results indicate that the cytochrome c oxidase biosensors could be used to determine variations in cytochrome c concentration and thus have potential to be used as a diagnostic tool in the detection of MIs and possibly also in the study of programmed cell death.
Ashe
Damian
D
Department of Preclinical Sciences, Biochemistry Unit, Faculty of Medical Sciences, University of the West Indies, St. Augustine, Eric Williams Medical Sciences Complex, Champs Fleurs, Trinidad and Tobago.
Alleyne
Trevor
T
Iwuoha
Emmanuel
E
D015374:Biosensing Techniques; D019411:Clinical Laboratory Techniques / Q000295:instrumentation*; D045304:Cytochromes c / Q000097:blood*; D003576:Electron Transport Complex IV / Q000378:metabolism; D006801:Humans; D008954:Models, Biological
D045304:Cytochromes c; D003576:Electron Transport Complex IV