Redox analysis of the cytochrome o-type quinol oxidase complex of Escherichia coli reveals three redox components.
Potentiometric analyses of the cytochrome o-type oxidase of Escherichia coli, using membranes from a strain containing amplified levels of the cytochrome bo complex, were conducted to resolve the redox centres of the oxidase. The cytochrome o-type oxidase of E. coli, a quinol oxidase, contains 2 mol of b-type haem per mol of complex and copper. Detailed analysis of potentiometric titrations, based on the absorbance of the Soret band, suggests that there are three contributions with midpoint potentials (Em,7) around +55 mV, +211 mV and +408 mV, all with maxima at 426-430 nm in the reduced state. In the alpha region of the spectra, a component with Em,6.85 = +58 mV has a maximal peak at 557 nm, and twin peaks at 556 and 564 nm nitrate with Em,6.85 = +227 mV. A feature corresponding to the highest potential Soret contribution was not observed. These data can be explained either by a model incorporating haem-haem interaction or by attributing the shorter-wavelength band (557 nm) to haem b and a split alpha-band (556, 564 nm) to the haem o (oxygen-binding haem b). Absolute spectra of oxidized membranes show continuous absorbance from 460 to 530 nm and suggest the presence of a high-spin haem component in the membranes. Monitoring absorbance at 635 minus 672 nm, contributions with midpoints (Em,7) around +52 mV, +234 mV and +371 mV are observed. This latter contribution is possibly the highest-potential component which titrates with Em greater than +400 mV in the Soret region and may represent copper-haem coupling in the cytochrome o complex.
Bolgiano
B
B
Microbial Physiology Research Group, Kings College London, U.K.
Salmon
I
I
Ingledew
W J
WJ
Poole
R K
RK
D004563:Electrochemistry; D004578:Electron Spin Resonance Spectroscopy; D003576:Electron Transport Complex IV / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology*; D010084:Oxidation-Reduction; D013057:Spectrum Analysis; D013696:Temperature
C038862:cytochrome o oxidase; D003576:Electron Transport Complex IV