PMID:11076519
| Citation |
Jasaitis, A, Borisov, VB, Belevich, NP, Morgan, JE, Konstantinov, AA and Verkhovsky, MI (2000) Electrogenic reactions of cytochrome bd. Biochemistry 39:13800-9 |
|---|---|
| Abstract |
Cytochrome bd is one of the two terminal quinol oxidases in the respiratory chain of Escherichia coli. The enzyme catalyzes charge separation across the bacterial membrane during the oxidation of quinols by dioxygen but does not pump protons. In this work, the reaction of cytochrome bd with O(2) and related reactions has been studied by time-resolved spectrophotometric and electrometric methods. Oxidation of the fully reduced enzyme by oxygen is accompanied by rapid generation of membrane potential (delta psi, negative inside the vesicles) that can be described by a two-step sequence of (i) an initial oxygen concentration-dependent, electrically silent, process (lag phase) corresponding to the formation of a ferrous oxy compound of heme d and (ii) a subsequent monoexponential electrogenic phase with a time constant <60 mus that matches the formation of ferryl-oxo heme d, the product of the reaction of O(2) with the 3-electron reduced enzyme. No evidence for generation of an intermediate analogous to the "peroxy" species of heme-copper oxidases could be obtained in either electrometric or spectrophotometric measurements of cytochrome bd oxidation or in a spectrophotometric study of the reaction of H(2)O(2) with the oxidized enzyme. Backflow of electrons upon flash photolysis of the singly reduced CO complex of cytochrome bd leads to transient generation of a delta psi of the opposite polarity (positive inside the vesicles) concurrent with electron flow from heme d to heme b(558) and backward. The amplitude of the delta psi produced by the backflow process, when normalized to the reaction yield, is close to that observed in the direct reaction during the reaction of fully reduced cytochrome bd with O(2) and is apparently associated with full transmembrane translocation of approximately one charge. |
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| Keywords |
Carbon Monoxide/chemistry; Cytochromes/chemistry; Electron Transport; Electron Transport Chain Complex Proteins; Escherichia coli/enzymology; Escherichia coli Proteins; Hydrogen Peroxide/chemistry; Membrane Potentials; Oxidation-Reduction; Oxidoreductases/chemistry; Oxygen/chemistry; Photolysis; Spectrophotometry |
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