SUIT-025 O2 mt D057: Difference between revisions
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{{MitoPedia | {{MitoPedia | ||
|abbr=FNS(Oct,PGM) | |abbr=FNS(Oct,PGM) | ||
|description=[[File:1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Rot;9Ama.png| | |description=[[File:1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Rot;9Ama.png|600px]] | ||
|info='''ย '''ย ย - '''[[SUIT-025]]''' | |info='''ย '''ย ย - '''[[SUIT-025]]''' | ||
|application=O2 | |application=O2 | ||
|SUIT number=D057_1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot;9Ama | |SUIT number=D057_1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot;9Ama | ||
}} | }} | ||
::: '''[[Categories of SUIT protocols|SUIT-category]]:''' FNS(Oct,PGM) | ::: '''[[Categories of SUIT protocols|SUIT-category]]:''' FNS(Oct,PGM) | ||
::: '''[[SUIT protocol pattern]]:''' lineal 1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot- | ::: '''[[SUIT protocol pattern]]:''' lineal 1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot- | ||
{{Template:SUIT text D057}} | |||
__TOC__ | __TOC__ | ||
ย Communicated by [[Doerrier C]] (last update 2019-05-21) | ย Communicated by [[Doerrier C]] (last update 2019-05-21) | ||
== Representative traces == | |||
[[File:D057 O2 traces.png|600px]] | |||
{{Template:SUIT-025}} | {{Template:SUIT-025}} | ||
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::::* [[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. | ::::* [[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. | ||
== Chemicals and syringes == | |||
{{Template:Chemicals SUIT-025}} | |||
::: Suggested stock concentrations are shown in the specific DL-Protocol. | |||
== References == | == References == |
Latest revision as of 10:29, 18 September 2020
Description
Abbreviation: FNS(Oct,PGM)
Reference: - SUIT-025
SUIT number: D057_1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot;9Ama
O2k-Application: O2
- SUIT-category: FNS(Oct,PGM)
- SUIT protocol pattern: lineal 1D;2M;3Oct;3c;4M;5P;6G;7S;8Rot-
The SUIT-025 O2 mt D057 protocol is based on SUIT-002 specially designed to give information on F-pathway in OXPHOS state avoiding fatty acid oxidation (FAO) overestimation in the presence of anaplerotic pathways. Moreover, the pathway control in OXPHOS state (F, F(N), FN, FNS, pathways) can be evaluated by using this SUIT protocol.
Communicated by Doerrier C (last update 2019-05-21)
Representative traces
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.
Chemicals and syringes
Step | Chemical(s) and link(s) | Comments |
---|---|---|
1D | ADP (D) | |
2M.1 | Malate (M) | |
3Oct | Octanoylcarnitine (Oct) | |
3c | Cytochrome c (c) | |
4M2 | Malate (M) | |
5P | Pyruvate (P) | |
6G | Glutamate (G) | |
7S | Succinate (S) | |
8Rot | Rotenone (Rot) | |
9Ama | Antimycin A (Ama) |
- Suggested stock concentrations are shown in the specific DL-Protocol.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry