SUIT-027: Difference between revisions
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:::- This protocol does not analyse [[Coupling control state|coupling control]]. | :::- This protocol does not analyse [[Coupling control state|coupling control]]. | ||
:::- Different [[pathway control states]] are not analysed in this protocol. | :::- Different [[pathway control states]] are not analysed in this protocol. | ||
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== References == | == References == | ||
{{#ask:[[Category:Publications]] [[Instrument and method::O2k-Protocol]] [[Additional label::SUIT- | {{#ask:[[Category:Publications]] [[Instrument and method::O2k-Protocol]] [[Additional label::SUIT-027]] | ||
|?Was published in year=Year | |?Was published in year=Year | ||
|?Has title=Reference | |?Has title=Reference |
Revision as of 08:52, 1 July 2019
Description
File:Ce1;1Dig;1M;2D;3M;4P;5G-.png
Abbreviation: Malate anaplerotic pathway
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 mitochondrial preparations (isolated mitochondria, permeabilized cells, homogenate and permeabilized fibers). Malate alone does not support respiration if the oxaloacetate is not metabolized further in the absence of acetyl-CoA. The careful titration of malate is required to analyse the activity of the mt-isoform of NADP+- or NAD(P)+-dependent malic enzyme (mtME). If these enzymes are present in the sample one should be able to reach a high respiratory activity comparable to the NADH linked pathway control states with more classical combinations of substrates (e.g. PM or GM).
Communicated by Cardoso LH (last update 2019-07-01)
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) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
1Dig | Dig | ce1;1Dig
|
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
|
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- 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 due to mitochondrial malic enzyme in the sample studied.
- Malate concentration that does not stimulate malate anaplerotic pathway, and therefore could be used to study F-pathway without N-pathway contribution in samples with mt malic enzyme.
- - This protocol does not analyse coupling control.
- - Different pathway control states are not analysed in this protocol.
- + Multiple titrations with malate provide information on:
Compare SUIT protocols
References
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry