Komlodi 2021 MitoFit Q

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Komlodi 2021 MitoFit Q

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Komlodi T, Cardoso LHD, Doerrier C, Gnaiger E (2021) Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria. MitoFit Preprints 2021.2. doi:10.26124/mitofit:2021-0002


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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, Gnaiger Erich (2021-02-18) MitoFit Prep

Abstract: Redox states of mitochondrial coenzyme Q (mtCoQ or Q) 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 oxygen, 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 the Q redox state continuously and simultaneously with oxygen consumption. The Q-Module was optimized for high signal-to-noise ratio and minimum oxygen diffusion. CoQ2 is added as a redox probe equilibrating with Q at Complexes CI, CII and CIII and the detecting electrode. Q-sensors are poised with the CoQ2 redox peak potentials determined by cyclic voltammetry, which provides quality control of the Q-sensor and reveals chemical interferences. The Q redox state and oxygen consumption were measured simultaneously in isolated mitochondria. A coupling-control protocol was applied to analyze LEAK, OXPHOS, and electron transfer capacities (L, P, and E, respectively) in the succinate-pathway. In a second pathway-control protocol, NADH- and succinate-linked pathways (N and S) converge at the Q-junction. mtCoQ was more oxidized when O2 flux was stimulated in coupling-control states with load increasing from L to P and E. In contrast, mtCoQ was more reduced when O2 flux was stimulated with electron input capacities increasing from N-, S- to NS-pathway-control states. N- and S- pathway capacities were not completely additive, thus confirming partial pool behavior of Q as proposed in the plasticity model of supercomplex organization.

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: ORCID.png Komlodi Timea, ORCID.png Cardoso Luiza HD, ORCID.png Doerrier Carolina, ORCID.png Gnaiger Erich


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Cited by

  • Huete-Ortega et al (2021) Substrate-uncoupler-inhibitor-titration protocols for photosynthesis in Chlamydomonas reinhardtii. MitoFit Preprints 2021 (in prep).

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