From Bioblast
Description
Abbreviation: FNS(Oct,GM)
Reference: A: SUIT-017
SUIT number: D046_1OctM;2D;3G;3c;4S;5U;6Rot;7Ama
O2k-Application: O2
MitoPedia concepts:
SUIT protocol,
SUIT A,
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MitoPedia methods:
Respirometry
- SUIT-category: FNS(Oct,GM)
- SUIT protocol pattern: diametral 1OctM;2D;3G;4S;5U;6Rot-
SUIT-017 O2 mt D046 gives information on F-pathway in LEAK state and OXPHOS state. In addition, the pathway control of FN and FNS in OXPHOS state and of FNS and S in ET state is evaluated.
Communicated by Cardoso LHD, Gnaiger E (last update 2019-07-31)
Representative traces
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1OctM | OctML(n) | F(N) | FAO | 1OctM
|
2D | OctMP | F(N) | FAO | 1OctM;2D
|
2c | OctMcP | FN | FAO | 1OctM;2D;2c
|
3G | OctGMP | FN | F&CI | 1OctM;2D;2c;3G;
|
4S | OctGMSP | FNS | F&CI&II | 1OctM;2D;2c;3G;4S
|
5U | OctGMSE | FNS | F&CI&II | 1OctM;2D;2c;3G;4S;5U
|
6Rot | SE | S | CII | 1OctM;2D;2c;3G;4S;5U;6Rot
|
7Ama | ROX | 1OctM;2D;2c;3G;4S;5U;6Rot;7Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
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- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- + The protocol provides information on F-OXPHOS capacity in the absence of a significant OXPHOS capacity in the malate-anaplerotic pathway control state.
- + Glutamate is easier to prepare compared to pyruvate.
- + Reasonable duration of the experiment.
- + This protocol can be extended with the Complex IV module, which can prolong the experimental time by ~30 min.
- - This protocol analyses only GM as NADH-linked substrates. Comparison of GM- with PM-capacity yields important information on N-pathway respiratory control upstream of CI (Lemieux et al 2017; Votion et al 2012).
- - F-OXPHOS capacity may be underestimated with Oct. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) & malate increased OXPHOS capacity by 26% (Lemieux et al 2011).
- + The protocol provides information on F-OXPHOS capacity in the absence of a significant OXPHOS capacity in the malate-anaplerotic pathway control state.
Compare SUIT protocols
Chemicals and syringes
Step | Chemical(s) and link(s) | Comments |
---|---|---|
1OctM | Octanoylcarnitine (Oct) and Malate (M) | |
2D | ADP (D) | |
2c | Cytochrome c (c) | |
3G | Glutamate (G) | |
4S | Succinate (S) | |
5U | Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U) | Can be substituted for other uncoupler |
6Rot | Rotenone (Rot) | |
7Ama | Antimycin A (Ama) |
- Suggested stock concentrations are shown in the specific DL-Protocol.