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Komlodi 2021 MitoFit Q

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Komlodi 2021 MitoFit Q

Publications in the MiPMap
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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MitoFit pdf

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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O2k-Manual
Mitochondrial Respiration Medium - MiR05-Kit.
Bioblast pdf Â» Versions2022-03-03
O2k-Manual
NextGen-O2k: Q-Module manual
PDF.jpg Â» Versions2021-10-29
O2k-Protocols
Laboratory protocol: isolation of mouse heart mitochondria.
Bioblast pdf Â»Versions2021-08-09

Keywords


Questions.jpg


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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


Cited by

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

Support

Template NextGen-O2k.jpg
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