Steps and respiratory states
[[File:ce1;ce2P;ce3Omy;ce4U;ce5Rot;ce6S;ce7Ama.png|300px]
Step
|
State
|
Pathway
|
Q-junction
|
Comment - Events (E) and Marks (M)
|
0DTPA
|
|
|
|
- DTPA is an iron chelator, which decreases the chemical fluorescence background created by the Amplex UltraRed assay. Administration of DTPA into the O2k-chamber is recommended before all other chemicals because the iron chelation capacity of the compound is time-dependent (approx. 10-15 min). However, the experiments can be carried out in the absence of DTPA.
|
0SOD
|
|
|
|
- SOD or superoxide dismutase converts the anion superoxide released by the mitochondria into H2O2, making it accessible to the Amplex UltraRed assay.
|
0HRP
|
|
|
|
- HRP or horseradish peroxidase catalyses the conversion of Amplex UltraRed and H2O2 towards the fluorescent resorufin.
|
0AmR
|
|
|
|
|
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.
|
ce1P
|
ROUTINE
|
|
|
ce1;ce1P; ROUTINE, R
- ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
|
ce2Omy
|
LOmy
|
|
|
ce1;ce1P;ce2Omy
- Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
|
ce3U
|
E
|
|
|
ce1;ce1P;ce2Omy;ce3U
|
ce4Rot
|
ROX
|
|
|
ce1;ce1P;ce2Omy;ce3U;ce4Rot
- 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.
|
ce5S
|
|
|
|
ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S
|
ce6Ama
|
|
|
|
ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;ce6Ama
- 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).
|