PMID: 9829979Nov 26, 1998

Chloride bound to oxidized cytochrome c oxidase controls the reaction with nitric oxide

The Journal of Biological Chemistry
Alessandro GiuffrèM T Wilson

Abstract

The reaction of nitric oxide (NO) with oxidized fast cytochrome c oxidase was investigated by stopped-flow, amperometry, and EPR, using the enzyme as prepared or after "pulsing." A rapid reduction of cytochrome a is observed with the pulsed, but not with the enzyme as prepared. The reactive species (lambdamax = 424 nm) reacts with NO at k = 2.2 x 10(5) M-1 s-1 at 20 degreesC and is stable for hours unless Cl- is added, in which case it decays slowly (t1/2 approximately 70 min) to an unreactive state (lambdamax = 423 nm) similar to the enzyme as prepared. Thus, Cl- binding prevents a rapid reaction of NO with the oxidized binuclear center. EPR experiments show no new signals within 15 s after addition of NO to the enzyme as prepared. Amperometric measurements show that the pulsed NO-reactive enzyme reacts with high affinity and a stoichiometry of 1 NO/aa3, whereas the enzyme as prepared reacts to a very small extent (<20%). In both cases, the reactivity is abolished by pre-incubation with cyanide. These experiments suggest that the effect of "pulsing" the enzyme, which leads to enhanced NO reactivity, arises from removing Cl- bound at the oxidized cytochrome a3-CuB site.

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Citations

Sep 5, 2002·Biochemical and Biophysical Research Communications·Clive ButlerPaolo Sarti
Mar 15, 2003·Clinica Chimica Acta; International Journal of Clinical Chemistry·Daniela GiustariniRanieri Rossi
Mar 5, 2003·Free Radical Biology & Medicine·Paolo SartiMaurizio Brunori
Feb 14, 2002·Trends in Biochemical Sciences·Chris E Cooper
Mar 28, 2012·Chemical Research in Toxicology·Jose Carlos Toledo, Ohara Augusto
Aug 2, 2007·Antioxidants & Redox Signaling·Fernando AntunesEnrique Cadenas
Jul 25, 2000·Biochemical and Biophysical Research Communications·Paolo SartiMaurizio Brunori
Jan 13, 2006·Proceedings of the National Academy of Sciences of the United States of America·Maria G MasonChristopher E Cooper
Apr 26, 2005·FEBS Letters·Alessandro GiuffrèPaolo Sarti
Feb 4, 2014·Biochimica Et Biophysica Acta·Alessandro GiuffrèElena Forte
May 16, 2015·Critical Reviews in Food Science and Nutrition·Catherine P BondonnoJonathan M Hodgson
Jun 24, 2006·Biochimica Et Biophysica Acta·Maurizio BrunoriPaolo Sarti
Apr 22, 2004·Biochimica Et Biophysica Acta·Marian FabianGraham Palmer
Apr 16, 2002·The Journal of Biological Chemistry·Alessandro GiuffrèPaolo Sarti
Feb 10, 2004·The Journal of Biological Chemistry·Marian FabianGraham Palmer
Apr 6, 2005·Biochemistry. Biokhimii︠a︡·S S KuznetsovaA A Konstantinov

Related Concepts

Bos indicus
Chloride Ion Level
Cytochrome c Oxidase Subunit VIa
Electrochemistry
Nitric Oxide
Oxidation-Reduction
Plasma Protein Binding Capacity

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