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Steps and respiratory states
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.
|
(ce2Omy)
|
L(Omy)
|
|
|
ce1;(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;(ce2Omy);ce3U
|
ce3Glc
|
E
|
|
|
ce1;(ce2Omy);ce3U;ce3Glc
|
ce3'U
|
E
|
|
|
ce1;(ce2Omy);ce3U;ce3Glc;ce3'U
- Noncoupled electron transfer state, ET state, with ET capacity E.
- Additional uncoupler titrations are performed to ensure reaching ET state after addition of glucose.
|
ce4Ama
|
ROX
|
|
|
ce1;(ce2Omy);ce3U;ce3Glc;ce3'U;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.
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- » Coupling control state
- » ET capacity
- » OXPHOS capacity
- » LEAK respiration
- Pathway control
- » Electron transfer pathway
- » Fatty acid oxidation pathway control state, F
- » NADH electron transfer-pathway state, N
- » Succinate pathway control state, S
- » NS-pathway control state, NS
- » Glycerophosphate pathway control state, Gp
- » Complex IV single step, CIV
- » Anaplerotic pathway control state
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » Succinate, S
- Glossary
- » List of SUIT states
- » SUIT concept