SUIT-027 O2 ce-pce D065: Difference between revisions
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{{MitoPedia concepts | {{MitoPedia concepts |
Revision as of 16:39, 27 June 2019
Description
Abbreviation: Anaplerotic pathway
Reference: A - SUIT-027
SUIT number: D065_ce1;1Dig;1M;2D;3M;4P;5G;6Ama
O2k-Application: O2
- MitoPedia: SUIT - Protocol for analysis of the anaplerotic pathway
- SUIT protocol pattern: ce1;1Dig;1M;2D;3M;4P;5G-
This protocol 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 our sample we 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 Iglesias-Gonzalez J and Gnaiger E (last update 2019-06-26)
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
- +
- -
Compare SUIT protocols
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Fluorometry