PMID:11551223
| Citation |
Léger, C, Heffron, K, Pershad, HR, Maklashina, E, Luna-Chavez, C, Cecchini, G, Ackrell, BA and Armstrong, FA (2001) Enzyme electrokinetics: energetics of succinate oxidation by fumarate reductase and succinate dehydrogenase. Biochemistry 40:11234-45 |
|---|---|
| Abstract |
Protein film voltammetry is used to probe the energetics of electron transfer and substrate binding at the active site of a respiratory flavoenzyme--the membrane-extrinsic catalytic domain of Escherichia coli fumarate reductase (FrdAB). The activity as a function of the electrochemical driving force is revealed in catalytic voltammograms, the shapes of which are interpreted using a Michaelis-Menten model that incorporates the potential dimension. Voltammetric experiments carried out at room temperature under turnover conditions reveal the reduction potentials of the FAD, the stability of the semiquinone, relevant protonation states, and pH-dependent succinate--enzyme binding constants for all three redox states of the FAD. Fast-scan experiments in the presence of substrate confirm the value of the two-electron reduction potential of the FAD and show that product release is not rate limiting. The sequence of binding and protonation events over the whole catalytic cycle is deduced. Importantly, comparisons are made with the electrocatalytic properties of SDH, the membrane-extrinsic catalytic domain of mitochondrial complex II. |
| Links | |
| Keywords |
Flavin-Adenine Dinucleotide; Flavoproteins/metabolism; Kinetics; Models, Chemical; Models, Theoretical; Oxidation-Reduction; Potentiometry/methods; Succinate Dehydrogenase/metabolism; Succinic Acid/metabolism; Thermodynamics |
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