- high-resolution terminology - matching measurements at high-resolution
SUIT-018
Description
Abbreviation: N(PGM)
Reference: A Β»Versions
- SUIT-category: N(PGM)
- SUIT protocol pattern: linear 1PGM;2D;3U
DLP applications
References
Steps and respiratory states
Step
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State
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Pathway
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Q-junction
|
Comment - Events (E) and Marks (M)
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0DTPA
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|
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- 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
|
|
|
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- 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)
|
1GMS
|
GMSL(n)
|
NS
|
CI&CII
|
1GMS
- Respiratory stimulation by simultaneous action of type N substrates & succinate, with convergent electron flow in the NS-pathway for reconstitution of TCA cycle function.
- Non-phosphorylating resting state (LEAK state); LEAK respiration L(n) in the absence of ADP, ATP, AMP (no adenylates).
|
2D
|
GMSP
|
NS
|
CI&CII
|
1GMS;2D
- Respiratory stimulation by simultaneous action of type N substrates & succinate, with convergent electron flow in the NS-pathway for reconstitution of TCA cycle function.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
3Ama
|
ROX
|
|
|
1GMS;2D;3Ama
- 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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Strengths and limitations
Compare SUIT protocols
MitoPedia concepts:
SUIT A
MitoPedia methods:
Respirometry
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