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.
|
ce1Snv
|
ROUTINE
|
|
|
ce1
- ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
- MitoKit-CII/Succinate-nv (diacetoxymethyl succinate) is a plasma membrane-permeable prodrug (permeable succinate; Snv) that diffuses across the plasma membrane. Cleavage of diacetoxymethyl residues is mediated by intracellular esterases, thus releasing succinate in the intracellular space. Abliva #: 01-118-s4
|
(ce2Omy)
|
L(Omy)
|
|
|
ce1;ce1Snv;(ce2Omy)
- Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
- The addition of oligomycin is optional depending on the experimental question to resolve.
|
ce3U
|
E
|
|
|
ce1;ce1Snv;(ce2Omy);ce3U
|
ce4Rot
|
ROX
|
|
|
ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot
- Rotenone inhibits Complex I, thus inhibiting the TCA cycle in living cells with intact plasma membrane. In the presence of Succinate-nv (diacetoxymethyl succinate, MitoKit-CII), the intracellular release of succinate independent of the TCA cycle leads to stimulation of respiration.
|
ce5Ama
|
ROX
|
|
|
ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot;ce5Ama
- 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.
|