Komlodi 2021 MitoFit Q

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Komlodi 2021 MitoFit Q

Komlódi T, Cardoso LHD, Doerrier C, Moore AL, Rich PR, Gnaiger E (2021) Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria. https://doi.org/10.26124/mitofit:2021-0002.v4 — 2021-11-11 published in Bioenerg Commun 2021.3.

» MitoFit Preprints 2021.2.v4.

Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria

MitoFit Preprints (2021-09-15) MitoFit Prep

Abstract: https://www.bioenergetics-communications.org/index.php/bec/article/view/komlodi_2021_amr Published in Bioenergetics Communications: 2021-11-11

Komlódi T, Cardoso LHD, Doerrier C, Moore AL, Rich PR, Gnaiger E (2021) Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria. Bioenerg Commun 2021.3. https://doi.org/10.26124/bec:2021-0003

Version 4 (v4) 2021-09-15 doi:10.26124/mitofit:2021-0002.v4

Version 3 (v3) 2021-09-01 doi:10.26124/mitofit:2021-0002.v3

Version 2 (v2) 2021-05-06 doi:10.26124/mitofit:2021-0002.v2

Version 1 (v1) 2021-02-18 doi:10.26124/mitofit:2021-0002 - »Link to all versions«

Redox states of the mitochondrial coenzyme Q pool, which reacts with the electron transfer system, reflect the balance between (1) reducing capacities of electron flow from fuel substrates converging at the Q-junction, (2) oxidative capacities downstream of Q to O₂, and (3) the load on the OXPHOS system utilizing or dissipating the protonmotive force.

A three-electrode sensor (Rich 1988; Moore et al 1988) was implemented into the NextGen-O2k to monitor continuously the redox state of CoQ2 added as a Q-mimetic simultaneously with O₂ consumption. The Q-Module was optimized for high signal-to-noise ratio, minimum drift, and minimum oxygen diffusion. CoQ2 equilibrates in the same manner as Q at Complexes CI, CII and CIII. The CoQ2 redox state is monitored amperometrically with the working electrode, which is poised at CoQ2 redox peak potentials determined by cyclic voltammetry. The voltammogram also provides quality control of the Q-sensor and reveals chemical interferences.

The CoQ2 redox state and O₂ consumption were measured simultaneously in isolated mouse cardiac and brain mitochondria. CoQ2 ― and by implication mitochondrial Q ― was more oxidized when O₂ flux was stimulated by coupling control: when energy demand increased from LEAK to OXPHOS and electron transfer capacities in the succinate pathway. In contrast, CoQ2 was more reduced when O₂ flux was stimulated by pathway-control of electron input capacities, increasing from the NADH (N)- to succinate (S)-linked pathway which converge at the Q-junction, with CI-Q-CIII and CII-Q-CIII segments, respectively. N- and S- respiratory pathway capacities were not completely additive, compatible with partitioning of Q intermediary between the solid-state and liquid-state models of supercomplex organization. The direct proportionality of CoQ2 reduction and electron input capacities through the CI-Q-CIII and CII-Q-CIII segments suggests that CoQ2 is accurately mimicking mitochondrial Q-redox changes. • Keywords: coenzyme Q; CoQ, Q-junction, Q-redox state, electron transfer system; ETS, ETS-reactive Q-pool; Q, mitochondrial coenzyme Q; mtCoQ, supercomplexed Q; free Q-pool according to the fluid-state model; Qfree, cyclic voltammetry; CV, high-resolution respirometry; HRR, isolated mitochondria; imt, mouse heart mitochondria, mouse brain mitochondria, oxygen consumption, SUIT protocols, coupling control, pathway control, NS-pathway, additivity • Bioblast editor: Komlodi T

ORCID: Komlodi Timea, Cardoso Luiza HD, Doerrier Carolina, Moore AL, Rich Peter R, Gnaiger Erich

Data availability

Original files are available Open Access at Zenodo repository: 10.5281/zenodo.4478400

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MiPNet22.10 MiR05-Kit	Mitochondrial Respiration Medium - MiR05-Kit.	2022-03-03	» Versions
MiPNet24.12 NextGen-O2k: Q-Module	NextGen-O2k: Q-Module manual	2021-10-29	» Versions
MiPNet20.06 IsolationMouseHeart-mt	Laboratory protocol: isolation of mouse heart mitochondria.	2021-08-09	»Versions

Keywords

Bioblast links: Q - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Coenzyme Q

» Coenzyme Q

» Quinone, Ubiquinone Q; oxidized

» Quinol, Ubiquinol QH₂; reduced

» Semiquinone

» Coenzyme Q2

» Q-redox state

» Q-pools

Mitochondrial pathways, respiratory Complexes, and Q

» Q-cycle

» Q-junction

» Convergent electron flow

» NS-pathway

» FNS

» FNSGp

» N-pathway

» Reverse electron flow from CII to CI

» CI

» Rotenone

» Amytal

» Piericidin

» S-pathway

» CII

» Malonate

» F-pathway

» CETF, Electron-transferring flavoprotein complex

» Gp-pathway

» CGpDH, Glycerophosphate dehydrogenase complex

» CIII

» Myxothiazol

» Choline dehydrogenase

» Dihydro-orotate dehydrogenase

NextGen-O2k and Q-Module

» NextGen-O2k

» Q-Module

» Q-Sensor

» Cyclic voltammetry

» Three-electrode system

General

» Categories of SUIT protocols

» Electron transfer pathway

» Electron-transfer-pathway state

» F-junction

» N-junction

Support

Supported by project NextGen-O2k which has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 859770.