Komlodi 2021 MitoFit Q
| Komlodi 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. MitoFit Preprints 2021.2.v2. doi:10.26124/mitofit:2021-0002.v2 |
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Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria
Komlodi Timea, Cardoso Luiza HD, Doerrier Carolina, Moore Anthony L, Rich Peter R, Gnaiger Erich (2021-05-06) MitoFit Prep
Abstract: Version 2 (v2) 2021-05-06 doi:10.26124/mitofit:190002.v2
- Version (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 CoQ ― was more oxidized when O2 flux was stimulated by coupling control: when load 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, thus confirming partial pool behavior 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: Q-junction, mitochondria, oxygen consumption, Q redox state, three-electrode system, cyclic voltammetry, harmonized SUIT protocols, high-resolution respirometry, coupling control, pathway control, NS-pathway, additivity • Bioblast editor: Komlodi T • O2k-Network Lab: AT Innsbruck Oroboros
ORCID: 
Komlodi Timea,
Cardoso Luiza HD,
Doerrier Carolina,
Rich Peter R,
Gnaiger Erich
Contents
References
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| MiPNet22.10 MiR05-Kit |  | » Versions | 2019-09-02 |
| MiPNet24.12 NextGen-O2k: Q-Module | | » Versions | 2021-01-27 |
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
- Coenzyme Q
- Mitochondrial pathways, respiratory Complexes, and Q
- » Q-cycle
- » Q-junction
- » Convergent electron flow
- » NS-pathway
- » FNS
- » FNSGp
- Mitochondrial pathways, respiratory Complexes, and Q
- NextGen-O2k and Q-Module
Cited by
- Huete-Ortega et al (2021) Substrate-uncoupler-inhibitor-titration protocols for photosynthesis in Chlamydomonas reinhardtii. MitoFit Preprints 2021 (in prep).
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.
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse
Tissue;cell: Heart, Nervous system
Preparation: Isolated mitochondria
Regulation: Redox state, Q-junction effect Coupling state: LEAK, OXPHOS, ET Pathway: N, S, NS HRR: Oxygraph-2k, NextGen-O2k
SUIT-006 Q mt D071, SUIT-006 Q ce-pce D073, SUIT-031 Q mt D072, SUIT-031 Q ce-pce D074, MitoFit 2021 Photosynthesis
