Rhodobacter capsulatus mutants lacking the Rieske FeS protein form a stable cytochrome bc1 subcomplex with an intact quinone reduction site.
The ubiquinol-cytochrome c oxidoreductase (or bc1 complex) of Rhodobacter capsulatus consists of three subunits: cytochrome b, cytochrome c1, and the Rieske iron-sulfur protein, encoded by the fbcF, fbcB, and fbcC genes, respectively. In the preceding paper [Davidson, E., Ohnishi, T., Atta-Asafo-Adjei, E., & Daldal, F. (1992) Biochemistry (preceding paper in this issue)], we have observed that the apoproteins for cytochromes b and c1 are fully present in the intracytoplasmic membrane of R. capsulatus mutants containing low amounts of, or no, Rieske apoprotein. Here we present evidence that the redox midpoint potentials of cytochromes b and c1, as well as their ability to bind antimycin and stabilize a semiquinone at the Qi site, are unaffected by the absence of the Rieske subunit. This is the first report describing a mutant containing a stable bc1 subcomplex with an intact Qi site in the chromatophore membranes, and provides further evidence that a functional quinone reduction site can be formed in the absence of a quinol oxidation (Qo) site. Additional mutants carrying fbc deletions expressing the remaining subunits of the cytochrome bc1 complex were constructed to investigate the relationship among these subunits for their stability in vivo. Western blot analysis of these mutants indicated that cytochromes b and c1 protect each other against degradation, suggesting that they form a two-protein subcomplex in the absence of the Rieske protein subunit.
Davidson
E
E
Department of Biology, School of Medicine, University of Pennsylvania, Philadelphia 19104.
Ohnishi
T
T
Tokito
M
M
Daldal
F
F
D000968:Antimycin A / Q000031:analogs & derivatives / Q000378:metabolism; D001665:Binding Sites; D015153:Blotting, Western; D014450:Electron Transport Complex III / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism; D016296:Mutagenesis; D010084:Oxidation-Reduction; D011809:Quinones / Q000378:metabolism*; D016354:Rhodobacter capsulatus / Q000737:chemistry* / Q000235:genetics; D013053:Spectrophotometry; D013329:Structure-Activity Relationship
D007506:Iron-Sulfur Proteins; D011809:Quinones; C032456:antimycin; D000968:Antimycin A; D014450:Electron Transport Complex III