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Template:SUIT-006 Q mt D071

From Bioblast
MitoPedia: SUIT

Steps and respiratory states

Mt;1Q2;1Rot;1S;2D;(3Omy);4U;5Anox;6Ama.png

Step State Pathway Q-junction Comment - Events (E) and Marks (M)
1Q2 REN 1Q2
  • To detect the redox state of the Q-pool a mimetic coenzyme Q2 is applied in suspension, which reacts both with the detecting electrode and the biological sites (CI, CII and CIII).
  • Respiration in the REN state is due to the presence of residual endogenous substrates in the sample preparation.
  • In the absence of endogenous substrates, this step can be used for calibration of fully oxidized Q.
1Rot ROX 1Rot
  • Rotenone is required to detect the fully oxidized Q-pool in the presence of isolated mitochondria (or permeabilized cells) and coenzyme Q2. Rotenone can inhibit the respiration supported by endogenous substrates that remained after the isolation procedure.
1S SL S CII 1Q2;1Rot;1S
2D SP S CII 1Q2;1Rot;1S;2D
(3Omy) SL(Omy) S CII 1Q2;1Rot;1S;2D;(3Omy)
  • Omy addition is skipped in SUIT-006 O2 mt D022
  • Succinate, S ( type S-pathway to Q).
  • Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
4U SE S CII 1Q2;1Rot;1S;2D;(3Omy);4U
5Anox S CII 1Q2;1Rot;1S;2D;(3Omy);4U;5Anox
  • Anoxia is a crucial step to detect the fully reduced Q-junction in the presence of coenzyme Q2 after the biological sample has consumed the O2 in the O2k-chamber for calculation of the Q redox state.
6Ama ROX 1Q2;1Rot;1S;2D;(3Omy);4U;5Anox;6Ama
  • Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated after addition of antimycin A (inhibitor of CIII). Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration (mt).
  • Since the previous step was anoxia, it is essential to reoxygenate the chamber before or after antimycin A titration to be able to measure Rox.


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