Photoinduced electron transfer between the Rieske iron-sulfur protein and cytochrome c(1) in the Rhodobacter sphaeroides cytochrome bc(1) complex - Effects of pH, temperature, and driving force
by Engstrom, Gregory; Xiao, Kunhong; Yu, Chang-An; Yu, Linda; Durham, Bill; Millett, Francis
Electron transfer from the Rieske iron-sulfur protein to cytochrome c(1) (cyt c(1)) in the Rhodobacter sphaeroides cytochrome bc(1) complex was studied using a ruthenium dimer complex, Ru2D. Laser flash photolysis of a solution containing reduced cyt bc(1), Ru2D, and a sacrificial electron acceptor results in oxidation of cyt c(1) within 1 mus, followed by electron transfer from the iron-sulfur center (2Fe-2S) to cyt c(1) with a rate constant of 80,000 s(-1). Experiments were carried out to evaluate whether the reaction was rate-limited by true electron transfer, proton gating, or conformational gating. The temperature dependence of the reaction yielded an enthalpy of activation of +17.6 kJ/mol, which is consistent with either rate-limiting conformational gating or electron transfer. The rate constant was nearly independent of pH over the range pH 7 to 9.5 where the redox potential of 2Fe-2S decreases significantly due to deprotonation of His-161. The rate constant was also not greatly affected by the Rieske iron-sulfur protein mutations Y156W, S154A, or S154A/Y156F, which decrease the redox potential of 2Fe-2S by 62, 109, and 159 mV, respectively. It is concluded that the electron transfer reaction from 2Fe-2S to cyt c(1) is controlled by conformational gating.
- Journal
- Journal of Biological Chemistry
- Volume
- 277
- Issue
- 34
- Year
- 2002
- Start Page
- 31072-31078
- URL
- https://dx.doi.org/10.1074/jbc.m202594200
- ISBN/ISSN
- 1083-351X; 0021-9258
- DOI
- 10.1074/jbc.m202594200