Characterization of the free energy dependence of an interprotein electron transfer reaction by variation of pH and site-directed mutagenesis.
| Autor: | Dow BA; Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL 32827, United States., Davidson VL; Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL 32827, United States. Electronic address: victor.davidson@ucf.edu. |
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| Jazyk: | angličtina |
| Zdroj: | Biochimica et biophysica acta [Biochim Biophys Acta] 2015 Oct; Vol. 1847 (10), pp. 1181-6. Date of Electronic Publication: 2015 Jun 15. |
| DOI: | 10.1016/j.bbabio.2015.06.012 |
| Abstrakt: | The interprotein electron transfer (ET) reactions of the cupredoxin amicyanin, which mediates ET from the tryptophan tryptophylquinone (TTQ) cofactor of methylamine dehydrogenase to cytochrome c-551i have been extensively studied. However, it was not possible to perform certain key experiments in that native system. This study examines the ET reaction from reduced amicyanin to an alternative electron acceptor, the diheme protein MauG. It was possible to vary the ΔG° for this ET reaction by simply changing pH to determine the dependence of kET on ΔG°. A P94A mutation of amicyanin significantly altered its oxidation-reduction midpoint potential value. It was not possible to study the ET from reduced P94A amicyanin to cytochrome c-551i in the native system because that reaction was kinetically coupled. However, the reaction from reduced P94A amicyanin to MauG was a true ET reaction and it was possible to determine values of reorganization energy (λ) and electronic coupling for the reactions of this variant as well as native amicyanin. Comparison of the λ values associated with the ET reactions between amicyanin and the TTQ of methylamine dehydrogenase, the diheme center of MauG and the single heme of cytochrome c-551i, provides insight into the factors that dictate the λ values for the respective reactions. These results demonstrate how study of ET reactions with alternative redox partner proteins can complement and enhance our understanding of the reactions with the natural redox partners, and further our understanding of mechanisms of protein ET reactions. (Copyright © 2015 Elsevier B.V. All rights reserved.) |
| Databáze: | MEDLINE |
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