Mutation of a single residue in the ba3 oxidase specifically impairs protonation of the pump site

Proceedings of the National Academy of Sciences of the United States of America
Christoph von BallmoosP Brzezinski

Abstract

The ba3-type cytochrome c oxidase from Thermus thermophilus is a membrane-bound protein complex that couples electron transfer to O2 to proton translocation across the membrane. To elucidate the mechanism of the redox-driven proton pumping, we investigated the kinetics of electron and proton transfer in a structural variant of the ba3 oxidase where a putative "pump site" was modified by replacement of Asp372 by Ile. In this structural variant, proton pumping was uncoupled from internal electron transfer and O2 reduction. The results from our studies show that proton uptake to the pump site (time constant ∼65 μs in the wild-type cytochrome c oxidase) was impaired in the Asp372Ile variant. Furthermore, a reaction step that in the wild-type cytochrome c oxidase is linked to simultaneous proton uptake and release with a time constant of ∼1.2 ms was slowed to ∼8.4 ms, and in Asp372Ile was only associated with proton uptake to the catalytic site. These data identify reaction steps that are associated with protonation and deprotonation of the pump site, and point to the area around Asp372 as the location of this site in the ba3 cytochrome c oxidase.

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Citations

Jul 1, 2016·Proceedings of the National Academy of Sciences of the United States of America·Ilsoo Kim, Arieh Warshel
Jul 20, 2016·Proceedings of the National Academy of Sciences of the United States of America·Camilla Rydström LundinPeter Brzezinski
Oct 1, 2016·International Journal of Molecular Sciences·Antonis NicolaidesConstantinos Varotsis
Sep 30, 2016·Scientific Reports·Simone GrafPeter Brzezinski
Mar 6, 2017·Biochimica Et Biophysica Acta. Bioenergetics·Catarina A Carvalheda, Andrei V Pisliakov
Jul 5, 2017·Scientific Reports·Rebecka AnderssonGisela Brändén
Jul 31, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Elena GorbikovaRuslan Kalendar
Jun 13, 2020·Biochimica Et Biophysica Acta. Bioenergetics·Xiuhong CaiM R Gunner
Nov 23, 2016·Biochimica Et Biophysica Acta. Bioenergetics·Markus L BjörckPeter Brzezinski
May 23, 2021·Biochimica Et Biophysica Acta. Bioenergetics·Sergey A SiletskyMårten Wikström
Jan 20, 2018·Chemical Reviews·Mårten WikströmVivek Sharma

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