SUIT-017 O2 mt D046: Difference between revisions
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|info='''A:''' '''[[SUIT-017]]''' | |info='''A:''' '''[[SUIT-017]]''' | ||
|application=O2 | |application=O2 | ||
|SUIT number=D046_1OctM;2D;3G;3c;4S;5U;6Rot;7Ama | |SUIT number=D046_1OctM;2D;2c;3G;3c;4S;5U;6Rot;7Ama | ||
}} | }} | ||
{{MitoPedia concepts | |||
|mitopedia concept=SUIT protocol, SUIT A, Find | |||
}} | |||
{{MitoPedia methods | |||
|mitopedia method=Respirometry | |||
}} | |||
{{MitoPedia O2k and high-resolution respirometry}} | |||
{{MitoPedia topics}} | |||
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' FNS(Oct,GM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' FNS(Oct,GM) | ||
::: '''[[SUIT protocol pattern]]:''' diametral 1OctM;2D;3G;4S;5U;6Rot | ::: '''[[SUIT protocol pattern]]:''' diametral 1OctM;2D;3G;4S;5U;6Rot- | ||
Β | |||
{{Template:SUIT text D046}} | |||
Β | |||
__TOC__ | __TOC__ | ||
Β Communicated by | Β Communicated by [[Cardoso LHD]], [[Gnaiger E]] (last update 2019-07-31) | ||
Β | |||
== Representative traces == | |||
Β | |||
[[File:D046 O2 traces.png|600px]] | |||
{{Template:SUIT-017 O2 mt D}} | {{Template:SUIT-017 O2 mt D}} | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::: | ::::+ The protocol provides information on F-OXPHOS capacity in the absence of a significant [[OXPHOS capacity| OXPHOS capacity]] in the [[malate-anaplerotic pathway control state]]. | ||
::::+ Glutamate is easier to prepare compared to pyruvate. | ::::+ Glutamate is easier to prepare compared to pyruvate. | ||
::::+ Reasonable duration of the experiment. | ::::+ Reasonable duration of the experiment. | ||
::::+ This protocol can be extended with the Complex IV module, which can prolong the experimental time | ::::+ This protocol can be extended with the Complex IV module, which can prolong the experimental time by ~30 min. | ||
::::- This protocol should not be used to analyze FAO in samples that express mitochondrial malic enzyme and thus present a significant [[OXPHOS capacity| OXPHOS capacity]] in the [[malate-anaplerotic pathway control state]]. | |||
::::- This protocol analyses only GM as NADH-linked substrates. Comparison of GM- with PM-capacity yields important information on N-pathway respiratory control upstream of CI (Lemieux et al 2017; Votion et al 2012). | |||
::::- F-OXPHOS capacity may be underestimated with Oct. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) & malate increased [[OXPHOS capacity| OXPHOS capacity]] by 26% ([[Lemieux_2011_Int_J_Biochem_Cell_Biol|Lemieux et al 2011]]). | |||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
::::* [[SUIT-002]] | ::::* [[SUIT-002]] or [[SUIT-025]] to measure F-OXPHOS capacity when malate anaplerotic activity is present; are also more comprehensive protocols including additional substrate states. | ||
::::* [[SUIT-005]] | ::::* [[SUIT-005]]: A comparable protocol using pyruvate (P) instead of glutamate (G). | ||
Β | |||
Β | |||
== Chemicals and syringes == | |||
{{Template:Chemicals SUIT-017}} | |||
::: Suggested stock concentrations are shown in the specific DL-Protocol. | |||
== References == | == References == | ||
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|sort=Was published in year | |sort=Was published in year | ||
|order=descending | |order=descending | ||
}} | }} |
Latest revision as of 11:45, 29 February 2024
Description
Abbreviation: FNS(Oct,GM)
Reference: A: SUIT-017
SUIT number: D046_1OctM;2D;2c;3G;3c;4S;5U;6Rot;7Ama
O2k-Application: O2
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry
- SUIT-category: FNS(Oct,GM)
- SUIT protocol pattern: diametral 1OctM;2D;3G;4S;5U;6Rot-
SUIT-017 O2 mt D046 gives information on F-pathway in LEAK state and OXPHOS state. In addition, the pathway control of FN and FNS in OXPHOS state and of FNS and S in ET state is evaluated.
Communicated by Cardoso LHD, Gnaiger E (last update 2019-07-31)
Representative traces
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1OctM | OctML(n) | F(N) | FAO | 1OctM
|
2D | OctMP | F(N) | FAO | 1OctM;2D
|
2c | OctMcP | FN | FAO | 1OctM;2D;2c
|
3G | OctGMP | FN | F&CI | 1OctM;2D;2c;3G;
|
4S | OctGMSP | FNS | F&CI&II | 1OctM;2D;2c;3G;4S
|
5U | OctGMSE | FNS | F&CI&II | 1OctM;2D;2c;3G;4S;5U
|
6Rot | SE | S | CII | 1OctM;2D;2c;3G;4S;5U;6Rot
|
7Ama | ROX | 1OctM;2D;2c;3G;4S;5U;6Rot;7Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- + The protocol provides information on F-OXPHOS capacity in the absence of a significant OXPHOS capacity in the malate-anaplerotic pathway control state.
- + Glutamate is easier to prepare compared to pyruvate.
- + Reasonable duration of the experiment.
- + This protocol can be extended with the Complex IV module, which can prolong the experimental time by ~30 min.
- - This protocol should not be used to analyze FAO in samples that express mitochondrial malic enzyme and thus present a significant OXPHOS capacity in the malate-anaplerotic pathway control state.
- - This protocol analyses only GM as NADH-linked substrates. Comparison of GM- with PM-capacity yields important information on N-pathway respiratory control upstream of CI (Lemieux et al 2017; Votion et al 2012).
- - F-OXPHOS capacity may be underestimated with Oct. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) & malate increased OXPHOS capacity by 26% (Lemieux et al 2011).
Compare SUIT protocols
Chemicals and syringes
Step | Chemical(s) and link(s) | Comments |
---|---|---|
1OctM | Octanoylcarnitine (Oct) and Malate (M) | |
2D | ADP (D) | |
2c | Cytochrome c (c) | |
3G | Glutamate (G) | |
4S | Succinate (S) | |
5U | Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U) | Can be substituted for other uncoupler |
6Rot | Rotenone (Rot) | |
7Ama | Antimycin A (Ama) |
- Suggested stock concentrations are shown in the specific DL-Protocol.