Movement of the Rieske iron-sulfur protein in the p-side bulk aqueous phase: effect of lumenal viscosity on redox reactions of the cytochrome b6f complex.
Based on the atomic structures of the mitochondrial cytochrome bc(1) complex, it has been proposed that the soluble domain of the [2Fe-2S] Rieske iron-sulfur protein (ISP) must rotate by ca. 60 degrees and translate through an appreciable distance between two binding sites, proximal to cytochrome c(1) and to the lumen-side quinol binding site. Such motional freedom implies that the electron-transfer rate should be affected by the lumenal viscosity. The flash-induced oxidation of cytochrome f, the chloroplast analogue of cytochrome c(1), was found to be inhibited reversibly by increased lumenal viscosity, as was the subsequent reduction of both cytochrome b(6) and cytochrome f. The rates of these three redox reactions correlated inversely with lumenal viscosity over a viscosity range of 1-10 cP. Reduction of cytochrome b(6) and cytochrome f was not concerted. The rate of cytochrome f reduction was observed to be approximately half that of cytochrome b(6) regardless of the actual viscosity, implying that the path length traversed by the ISP in reduction of cytochrome f is twice that of cytochrome b(6). This suggests that upon initiation of electron transfer by a light flash, cytochrome b(6) reduction requires movement of reduced ISP from an initial position predominantly proximal to cytochrome f, apparently favored by the reduced ISP, to the quinol binding site at which the oxidant-induced reduction of cytochrome b(6) is initiated. Subsequent reduction of cytochrome f requires the additional movement of the ISP back to a site proximal to cytochromef. There is no discernible viscosity dependence for cytochrome b(6) reduction under oxidizing conditions, presumably because the oxidized ISP preferentially binds proximal to the quinone binding niche. The dependence of the cytochrome redox reaction on ambient viscosity implies that the tethered diffusional motion of the ISP is part of the rate limitation for charge transfer through the b(6)f complex.
Heimann
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S
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
Ponamarev
M V
MV
Cramer
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WA
D002736:Chloroplasts / Q000737:chemistry / Q000378:metabolism; D003573:Cytochrome b Group / Q000737:chemistry* / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D014450:Electron Transport Complex III; D005947:Glucose / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry; D007202:Indicators and Reagents; D007506:Iron-Sulfur Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D009038:Motion; D010084:Oxidation-Reduction; D010971:Plastoquinone / Q000031:analogs & derivatives / Q000737:chemistry; D012995:Solubility; D018724:Spinacia oleracea; D013395:Sucrose / Q000737:chemistry; D014783:Viscosity; D014867:Water
D003573:Cytochrome b Group; D007202:Indicators and Reagents; D007506:Iron-Sulfur Proteins; D046911:Macromolecular Substances; C037989:Rieske iron-sulfur protein; D014867:Water; C003165:plastoquinol; D013395:Sucrose; D045346:Cytochrome b6f Complex; D014450:Electron Transport Complex III; D005947:Glucose; D010971:Plastoquinone; D005990:Glycerol