SUIT-025: Difference between revisions
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ย Communicated by [[Doerrier C]] (last update 2019-05- | ย Communicated by [[Doerrier C]] (last update 2019-05-21) | ||
== Specific SUIT protocols == | == Specific SUIT protocols == |
Revision as of 17:40, 21 May 2019
Description
Abbreviation: FNS(Oct,PGM)
Reference: ยปVersions
- SUIT-category: FNS(Oct,PGM)
- SUIT protocol pattern: lineal 1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot-
The SUIT-025 protocol is based on SUIT-002 specially designed to give information on F-pathway in OXPHOS state avoiding FAO overestimation in the presence of anaplerotic pathways. Moreover, the ET-pathway control of F, F(N), FN and FNS pathways in OXPHOS state can be evaluated by using this SUIT protocol.
Communicated by Doerrier C (last update 2019-05-21)
Specific SUIT protocols
SUIT-025 O2 mt D057
- SUIT-025 O2 mt D057 for isolated mitochondria, tissue homogenate and permeabilized cells (already permeabilized when they are added to the chamber)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1D | ROX | 1D
| ||
2M.1 | ||||
3Oct | OctMP | F | FAO | 1D;2M.1;3Oct
|
3c | OctMcP | F | FAO | 1D;2M.1;3Oct;3c
|
4M2 | OctMP | F(N) | FAO | 1D;2M.1;3Oct;4M2
|
5P | OctPMP | FN | FAO&CI | 1D;2M.1;3Oct;4M2;5P
|
6G | OctPGMP | FN | FAO&CI | 1D;2M.1;3Oct;4M2;5P;6G
|
7S | OctPGMSP | FNS | FAO&CI&II | 1D;2M.1;3Oct;4M2;5P;6G;7S
|
8Rot | SP | S | CII | 1D;2M.1;3Oct;4M2;5P;6G;7S;8Rot
|
9Ama | ROX | 1D;2M.1;3Oct;4M2;5P;6G;7S;8Rot;9Ama
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- ยป Electron transfer pathway
- ยป Fatty acid oxidation pathway control state, F
- ยป NADH electron transfer-pathway state, N
- ยป Succinate pathway control state, S
- ยป NS-pathway control state, NS
- ยป Glycerophosphate pathway control state, Gp
- ยป Complex IV single step, CIV
- ยป Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- ยป Glutamate, G
- ยป Glycerophosphate, Gp
- ยป Malate, M
- ยป Octanoylcarnitine, Oct
- ยป Pyruvate, P
- ยป Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- + SUIT-025 allows the depletion of endogenous substrates with ADP (1D).
- + The protocol provides information on F-pathway in OXPHOS state. The low concentration of malate used in this protocol, typically 0.1 mM, does not saturate the N-pathway; but saturates the F-pathway.
- + F-pathway (3Oct-2M.1) can be compared to FN-pathway (5P) in OXPHOS state.
- + Pathway control in OXPHOS (F, F(N), FN, FNS and S pathways) can be evaluated.
- + Harmonization with many SUIT protocols (up to step 7S).
- + Short experimental time.
- - ET state is not evaluated (for obtaining ET capacity with FNSGp and SGp substrate combinations see SUIT-002). To evaluate limitation of OXPHOS capacity by the phosphorylation system is it suggested to performed in parallel the SUIT-006 O2 mt D022 or to titrate the uncoupler after rotenone.
Compare SUIT protocols
- SUIT-002 (the reference protocol 2) differs from SUIT-025 in the coupling and pathway control states. SUIT-002 allows the evaluation of pathway control in OXPHOS state (F, F(N), FN, FNS, FNSGp pathways) and in ET state (FNSGp and SGp) whereas in SUIT-025 only F, F(N), FN and FNS pathways in OXPHOS state can be evaluated.
References
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry