SUIT-027: Difference between revisions
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::: '''[[SUIT protocol pattern]]:''' ce1;1Dig;1M;2D;3M;4P;5G | ::: '''[[SUIT protocol pattern]]:''' ce1;1Dig;1M;2D;3M;4P;5G | ||
SUIT-027 is focused on the analysis of the [[Malate anaplerotic pathway control state|malate anaplerotic pathway control state]] in [[mitochondrial preparations|mt-preparations]] (isolated mitochondria, tissue homogenate, permeabilized cells and permeabilized fibers). [[Malate]] alone does not support respiration of mt-preparations if oxaloacetate cannot be metabolized further in the absence of a source of acetyl-CoA. However, if malate anaplerotic pathway control state is present (e.g. [[Malic enzyme|mt-malic enzyme | SUIT-027 is focused on the analysis of the [[Malate anaplerotic pathway control state|malate anaplerotic pathway control state]] in [[mitochondrial preparations|mt-preparations]] (isolated mitochondria, tissue homogenate, permeabilized cells and permeabilized fibers). [[Malate]] alone does not support respiration of mt-preparations if oxaloacetate cannot be metabolized further in the absence of a source of acetyl-CoA. However, if malate anaplerotic pathway control state is present (e.g. [[Malic enzyme|mt-malic enzyme, mtME]]), then malate alone supports mt-respiration. SUIT-027 includes a careful titration of malate which is required to analyze the malate anaplerotic pathway control state. | ||
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Revision as of 10:04, 30 April 2020
Description
Abbreviation: Malate anaplerosis
Reference: A: Malate anaplerotic pathway
- SUIT protocol pattern: ce1;1Dig;1M;2D;3M;4P;5G
SUIT-027 is focused on the analysis of the malate anaplerotic pathway control state in mt-preparations (isolated mitochondria, tissue homogenate, permeabilized cells and permeabilized fibers). Malate alone does not support respiration of mt-preparations if oxaloacetate cannot be metabolized further in the absence of a source of acetyl-CoA. However, if malate anaplerotic pathway control state is present (e.g. mt-malic enzyme, mtME), then malate alone supports mt-respiration. SUIT-027 includes a careful titration of malate which is required to analyze the malate anaplerotic pathway control state.
Communicated by Cardoso LHD, Garcia-Souza LF and Doerrier C (last update 2020-04-30)
Specific SUIT protocols
- SUIT-027 O2 ce-pce D065 for permeabilized cells
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1M.05 | (ML) | ce1;1Dig;1M | ||
2D | ( MP) | ce1;1Dig;1M;2D
| ||
3M | MP | N | CI | ce1;1Dig;1M;2D;3M
|
4P | PMP | N | CI | ce1;1Dig;1M;2D;3M;4P
|
5G | PGMP | N | CI | ce1;1Dig;1M;2D;3M;4P;5G
|
6Ama | ROX | ce1;1Dig;1M;2D;3M;4P;5G;6Ama
|
- 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
- + Multiple titrations with malate provide information on:
- The presence of malate anaplerotic pathway (e.g. mt-malic enzyme) in the studied sample.
- Malate concentration which does not stimulate malate anaplerotic pathway, and therefore could be used to study F-pathway in OXPHOS state avoiding FAO overestimation in the presence of anaplerotic pathways (see SUIT-002).
- - This SUIT protocol does not analyze coupling control.
- - Different pathway control states are not analyzed in this SUIT protocol.
- + Multiple titrations with malate provide information on:
Compare SUIT protocols
- SUIT-007: SUIT protocol to analyze glutamate anaplerotic pathway.
- SUIT-002: 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-027 only malate anaplerotic pathway and N-pathway in OXPHOS state can be evaluated. A low concentration of malate is initially used to assess F-pathway without N-pathway contribution. Further, a higher concentration of malate is added to analyze FN-pathway.
- SUIT-025: SUIT-025 allows the evaluation of different ET-pathway states (F, F(N), FN, FNS and S) in OXPHOS state. A low concentration of malate is initially used to assess F-pathway without N-pathway contribution. Further, a higher concentration of malate is added to analyze FN-pathway.
References
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry