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SUIT-005
Description
Abbreviation: FNS(Oct,PM)
Reference: A: short RP2 Β»Versions
- SUIT-category: FNS(Oct,PM)
- SUIT protocol pattern: orthogonal 1OctM;2D;3P;4S;5U;6Rot;7Ama
Communicated by Iglesias-Gonzalez J, Doerrier C and Gnaiger E (last update 2019-01-21)
DLP applications
References
Steps and respiratory states
Step
|
State
|
Pathway
|
Q-junction
|
Comment - Events (E) and Marks (M)
|
1OctM
|
OctML(n)
|
F(N)
|
FAO
|
1OctM
- Respiratory stimulation of the FAO-pathway, F, by fatty acid FA in the presence of malate M. Malate is a type N substrate (N), required for the F-pathway. In the presence of anaplerotic pathways (e.g., mitochondrial malic enzyme, mtME) the F-pathway capacity is overestimated, if there is an added contribution of NADH-linked respiration, F(N) (see SUIT-002). The FA concentration has to be optimized to saturate the FAO-pathway, without inhibiting or uncoupling respiration.
- Low concentration of malate, typically 0.1 mM, does not saturate the N-pathway; but saturates the F-pathway.
- Non-phosphorylating resting state (LEAK state); LEAK respiration L(n) in the absence of ADP, ATP, AMP (no adenylates).
|
2D
|
OctMP
|
F(N)
|
FAO
|
1OCtM;2D
- Respiratory stimulation of the FAO-pathway, F, by fatty acid FA in the presence of malate M. Malate is a type N substrate (N), required for the F-pathway. In the presence of anaplerotic pathways (e.g., mitochondrial malic enzyme, mtME) the F-pathway capacity is overestimated, if there is an added contribution of NADH-linked respiration, F(N) (see SUIT-002). The FA concentration has to be optimized to saturate the FAO-pathway, without inhibiting or uncoupling respiration.
- Low concentration of malate, typically 0.1 mM, does not saturate the N-pathway; but saturates the F-pathway.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
2c
|
OctMcP
|
F(N)
|
FAO
|
1OCtM;2D;2c
- Respiratory stimulation of the FAO-pathway, F, by fatty acid FA in the presence of malate M. Malate is a type N substrate (N), required for the F-pathway. In the presence of anaplerotic pathways (e.g., mitochondrial malic enzyme, mtME) the F-pathway capacity is overestimated, if there is an added contribution of NADH-linked respiration, F(N) (see SUIT-002). The FA concentration has to be optimized to saturate the FAO-pathway, without inhibiting or uncoupling respiration.
- Low concentration of malate, typically 0.1 mM, does not saturate the N-pathway; but saturates the F-pathway.
- Addition of cytochrome c yields a test for integrity of the mtOM (cytochrome c control efficiency). Stimulation by added cytochrome c would indicate an injury of the mtOM and limitation of respiration in the preceding state without added c due to loss of cytochrome c. Typically, cytochrome c is added immediately after the earliest ADP-activation step (OXPHOS capacity P with saturating [ADP]).
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
3P
|
OctPMP
|
FN
|
F&CI
|
1OctM;2D;2c;3P
- Respiratory stimulation by simultaneous action of the F-pathway and N-pathway with convergent electron flow in the FN-pathway for evaluation of an additive or inhibitory effect of F.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
4S
|
OctPMSP
|
FNS
|
F&CI&II
|
1OctM;2D;2c;3P;4S
- Respiratory stimulation by simultaneous action of the F-pathway, N-pathway, and S-pathway, with convergent electron flow in the FNS-pathway for reconstitution of TCA cycle function and additive or inhibitory effect of F.
- OXPHOS capacity P (with saturating [ADP]), active OXPHOS state.
|
5U
|
OctPMSE
|
FNS
|
F&CI&II
|
1OctM;2D;2c;3P;4S;5U
|
6Rot
|
SE
|
S
|
CII
|
1OctM;2D;3P;4S;5U;6Rot
|
7Ama
|
ROX
|
|
|
1OctM;2D;3P;4S;5U;6Rot;7Ama
- 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.
|
Step
|
Respiratory state
|
Pathway control
|
ET-Complex
|
Comment
|
## AsTm
|
AsTmE
|
CIV
|
CIV
|
|
## Azd
|
CHB
|
|
|
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Β» Coupling control state
- Β» ET capacity
- Β» OXPHOS capacity
- Β» LEAK respiration
- 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
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Glossary
- Β» List of SUIT states
- Β» SUIT concept
Strengths and limitations
Compare SUIT protocols
MitoPedia concepts:
MiP concept,
SUIT protocol,
Recommended
MitoPedia methods:
Respirometry
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