Step
|
State
|
Pathway
|
Q-junction
|
Comment - Events (E) and Marks (M)
|
ce1
|
ROUTINE
|
|
|
ce1
- ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
|
ce1Glc
|
ROUTINE
|
|
|
ce1;ce1Glc
- ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
- Glucose is added to test Crabtree effect.
|
(ce2Omy)
|
L(Omy)
|
|
|
ce1;ce1Glc;(ce2Omy)
- Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
- If the aim is to analyse only Crabtree effect, it is recommended to skip this step. However, if there is also interest in studying coupling control, Omy can be added at this point to reach LEAK.
|
ce3U
|
E
|
|
|
ce1;ce1Glc;(ce2Omy);ce3U
|
ce4Ama
|
ROX
|
|
|
ce1;ce1Glc;(ce2Omy);ce3U;ce4Ama
- Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated either after inhibition of CIII (e.g. antimycin A, myxothiazol), CIV (e.g. Cyanide) or in the absence of endogenous fuel-substrates. Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration.
|