Time dependence of the catalytic intermediates in cytochrome c oxidase

Sanghwa Han, Satoshi Takahashi, Denis L. Rousseau

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Cytochrome c oxidase, the terminal enzyme in the electron transfer chain, catalyzes the reduction of oxygen to water in a multiple step process by utilizing four electrons from cytochrome c. To study the reaction mechanism, the resonance Raman spectra of the intermediate states were measured during single turnover of the enzyme after catalytic initiation by photolysis of CO from the fully reduced CO-bound enzyme. By measuring the change in intensity of lines associated with heme a, the electron transfer steps were determined and found to be biphasic with apparent rate constants of ~40 x 103 s-1 and ~1 x 103 s-1. The time dependence for the oxidation of heme a and for the measured formation and decay of the oxy, the ferryl ('F'), and the hydroxy intermediates could be simulated by a simple reaction scheme. In this scheme, the presence of the 'peroxy' ('P') intermediate does not build up a sufficient population to be detected because its decay rate is too fast in buffered H2O at neutral pH. A comparison of the change in the spin equilibrium with the formation of the hydroxy intermediate demonstrates that this intermediate is high spin. We also confirm the presence of an oxygen isotopesensitive line at 355 cm-1, detectable in the spectrum from 130 to 980 μs, coincident with the presence of the F intermediate.

Original languageEnglish (US)
Pages (from-to)1910-1919
Number of pages10
JournalJournal of Biological Chemistry
Volume275
Issue number3
DOIs
StatePublished - Jan 21 2000

Fingerprint

Electron Transport Complex IV
Electrons
Carbon Monoxide
Enzymes
Oxygen
Photolysis
Cytochromes c
Raman scattering
Rate constants
Oxidation
Water
Population
heme a

ASJC Scopus subject areas

  • Biochemistry

Cite this

Time dependence of the catalytic intermediates in cytochrome c oxidase. / Han, Sanghwa; Takahashi, Satoshi; Rousseau, Denis L.

In: Journal of Biological Chemistry, Vol. 275, No. 3, 21.01.2000, p. 1910-1919.

Research output: Contribution to journalArticle

Han, Sanghwa ; Takahashi, Satoshi ; Rousseau, Denis L. / Time dependence of the catalytic intermediates in cytochrome c oxidase. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 3. pp. 1910-1919.
@article{1caeeb2273e44de7adfd6a3b18fd3c96,
title = "Time dependence of the catalytic intermediates in cytochrome c oxidase",
abstract = "Cytochrome c oxidase, the terminal enzyme in the electron transfer chain, catalyzes the reduction of oxygen to water in a multiple step process by utilizing four electrons from cytochrome c. To study the reaction mechanism, the resonance Raman spectra of the intermediate states were measured during single turnover of the enzyme after catalytic initiation by photolysis of CO from the fully reduced CO-bound enzyme. By measuring the change in intensity of lines associated with heme a, the electron transfer steps were determined and found to be biphasic with apparent rate constants of ~40 x 103 s-1 and ~1 x 103 s-1. The time dependence for the oxidation of heme a and for the measured formation and decay of the oxy, the ferryl ('F'), and the hydroxy intermediates could be simulated by a simple reaction scheme. In this scheme, the presence of the 'peroxy' ('P') intermediate does not build up a sufficient population to be detected because its decay rate is too fast in buffered H2O at neutral pH. A comparison of the change in the spin equilibrium with the formation of the hydroxy intermediate demonstrates that this intermediate is high spin. We also confirm the presence of an oxygen isotopesensitive line at 355 cm-1, detectable in the spectrum from 130 to 980 μs, coincident with the presence of the F intermediate.",
author = "Sanghwa Han and Satoshi Takahashi and Rousseau, {Denis L.}",
year = "2000",
month = "1",
day = "21",
doi = "10.1074/jbc.275.3.1910",
language = "English (US)",
volume = "275",
pages = "1910--1919",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Time dependence of the catalytic intermediates in cytochrome c oxidase

AU - Han, Sanghwa

AU - Takahashi, Satoshi

AU - Rousseau, Denis L.

PY - 2000/1/21

Y1 - 2000/1/21

N2 - Cytochrome c oxidase, the terminal enzyme in the electron transfer chain, catalyzes the reduction of oxygen to water in a multiple step process by utilizing four electrons from cytochrome c. To study the reaction mechanism, the resonance Raman spectra of the intermediate states were measured during single turnover of the enzyme after catalytic initiation by photolysis of CO from the fully reduced CO-bound enzyme. By measuring the change in intensity of lines associated with heme a, the electron transfer steps were determined and found to be biphasic with apparent rate constants of ~40 x 103 s-1 and ~1 x 103 s-1. The time dependence for the oxidation of heme a and for the measured formation and decay of the oxy, the ferryl ('F'), and the hydroxy intermediates could be simulated by a simple reaction scheme. In this scheme, the presence of the 'peroxy' ('P') intermediate does not build up a sufficient population to be detected because its decay rate is too fast in buffered H2O at neutral pH. A comparison of the change in the spin equilibrium with the formation of the hydroxy intermediate demonstrates that this intermediate is high spin. We also confirm the presence of an oxygen isotopesensitive line at 355 cm-1, detectable in the spectrum from 130 to 980 μs, coincident with the presence of the F intermediate.

AB - Cytochrome c oxidase, the terminal enzyme in the electron transfer chain, catalyzes the reduction of oxygen to water in a multiple step process by utilizing four electrons from cytochrome c. To study the reaction mechanism, the resonance Raman spectra of the intermediate states were measured during single turnover of the enzyme after catalytic initiation by photolysis of CO from the fully reduced CO-bound enzyme. By measuring the change in intensity of lines associated with heme a, the electron transfer steps were determined and found to be biphasic with apparent rate constants of ~40 x 103 s-1 and ~1 x 103 s-1. The time dependence for the oxidation of heme a and for the measured formation and decay of the oxy, the ferryl ('F'), and the hydroxy intermediates could be simulated by a simple reaction scheme. In this scheme, the presence of the 'peroxy' ('P') intermediate does not build up a sufficient population to be detected because its decay rate is too fast in buffered H2O at neutral pH. A comparison of the change in the spin equilibrium with the formation of the hydroxy intermediate demonstrates that this intermediate is high spin. We also confirm the presence of an oxygen isotopesensitive line at 355 cm-1, detectable in the spectrum from 130 to 980 μs, coincident with the presence of the F intermediate.

UR - http://www.scopus.com/inward/record.url?scp=0034695481&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034695481&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.3.1910

DO - 10.1074/jbc.275.3.1910

M3 - Article

C2 - 10636892

AN - SCOPUS:0034695481

VL - 275

SP - 1910

EP - 1919

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -