Electron transfer in Paracoccus denitrificans with the modified fbc operon.
Membrane fragments of two mutant strains of Paracoccus denitrificans genetically modified in the bc(1) complex have been studied for comparison of enzymic activities of succinate-cytochrome-c reductase and its components, viz. succinate dehydrogenase (Complex II) and ubiquinol-cytochrome-c reductase (Complex III) and their response to changes in concentration of succinate, cytochrome c, ionic strength, pH, temperature and sensitivity to antimycin A. The mutants synthesized and assembled the b and c hemes in the ratio characteristic for the wild type strain. The mutant strain M 71 expressing the truncated copy of cytochrome c(1) (devoid of a stretch of 150 mainly acidic amino acids) was less sensitive to increasing concentration of cytochrome c and changes in ionic strength of the medium, but maintained the original affinity to succinate and sensitivity to antimycin A. The mutant strain M 36 with an overexpressed bc(1) content showed the highest response to changes in ionic strength and physical parameters, exhibited the lowest turnover number values with succinate-cytochrome-c reductase, but positively affected the succinate dehydrogenase. In view of the interaction of the redox components in native membranes the functional analyses of separated Complexes II and III should be regarded with caution.
Dadák
V
V
Institute of Biochemistry, Faculty of Science, Masaryk University, Brno, Czechia. dadak@chemi.muni.cz
Dudák
J
J
Zboril
P
P
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000968:Antimycin A / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D014450:Electron Transport Complex III; D004583:Electrons; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D009876:Operon; D009997:Osmotic Pressure; D010084:Oxidation-Reduction; D010231:Paracoccus denitrificans / Q000378:metabolism*; D013312:Stress, Physiological; D013384:Succinate Cytochrome c Oxidoreductase; D013696:Temperature
D000900:Anti-Bacterial Agents; D000968:Antimycin A; D013384:Succinate Cytochrome c Oxidoreductase; D014450:Electron Transport Complex III