Difference between revisions of "SUIT-027"
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{{MitoPedia | {{MitoPedia | ||
|abbr=[[Malate anaplerotic pathway control state|Malate | |abbr= [[Malate-anaplerotic pathway control state|Malate anaplerosis]] | ||
|description=[[File:1M;2D;3M;4P;5G-.png|400px]] | |description=[[File:1M;2D;3M;4P;5G-.png|400px]] | ||
|info='''A:''' '''[[Malate anaplerotic pathway control state|Malate anaplerotic pathway]]''' | |info='''A:''' '''[[Malate-anaplerotic pathway control state|Malate anaplerotic pathway]]''' | ||
}} | }} | ||
::: '''[[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, 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. | |||
__TOC__ | __TOC__ | ||
Communicated by [[Cardoso | Communicated by [[Cardoso LHD]], [[Garcia-Souza LF]] and [[Doerrier Carolina|Doerrier C]] (last update 2020-04-30) | ||
== Specific SUIT protocols == | == Specific SUIT protocols == | ||
Line 21: | Line 20: | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::+ Multiple titrations with [[malate]] provide information on: | :::+ Multiple titrations with [[malate]] provide information on: | ||
:::# The presence of malate anaplerotic pathway | :::# The presence of malate anaplerotic pathway (e.g. [[malic enzyme|mt-malic enzyme]]) in the studied sample. | ||
:::# Malate concentration | :::# 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 protocol does not | :::- This SUIT protocol does not analyze [[Coupling control state|coupling control]]. | ||
:::- Different [[pathway control states]] are not | :::- Different [[pathway control states]] are not analyzed in this SUIT protocol. | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
::::* [[SUIT-007]]: | ::::* [[SUIT-007]]: SUIT protocol to analyze glutamate anaplerotic pathway. | ||
::::* [[SUIT-002]]: 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 | ::::* [[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 == | == References == |
Latest revision as of 12:04, 3 June 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
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » 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