Step
|
State
|
Pathway
|
Q-junction
|
Comment - Events (E) and Marks (M)
|
1X
|
XL
|
|
|
ce1;Dig1;1X
- Electron-transfer-pathway state will depend on the substrate/inhibitor combinations added to fuel the mitochondrial respiration
- Non-phosphorylating resting state (LEAK state); LEAK respiration L(n) in the absence of ADP, ATP, AMP (no adenylates).
|
2D
|
XP
|
|
|
ce1;Dig1;1X;2D
|
2c
|
XcP
|
|
|
ce1;Dig1;1X;2D;2c
- Electron-transfer-pathway state will depend on the substrate/inhibitor combinations added to fuel the mitochondrial respiration
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
- Addition of cytochrome c yields a test for integrity of the mtOM (cytochrome c control efficiency). Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in the preceding state without added c due to loss of cytochrome c. Typically, cytochrome c is added immediately after the earliest ADP-activation step (OXPHOS capacity P with saturating [ADP]).
|
3(Omy)
|
XL
|
|
|
ce1;Dig1;1X;2D;2c;3(Omy)
- Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
- The addition of oligomycin or its carrier (EtOH) is optional depending on the experimental question to resolve. If we add only the carrier we do not reach LEAK state and we will evaluate if the EtOH alters the ROUTINE state.
|
4U
|
XE
|
|
|
ce1;Dig1;1X;2D;2c;3(Omy);4U
|
5*
|
ROX
|
|
|
ce1;Dig1;1X;2D;2c;3(Omy);4U;5*
- (*) Rox obtaining after inhibiting the CIII (e.g. antimycin A, myxothiazol).
- 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).
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