SUIT-005: Difference between revisions
From Bioblast
No edit summary |
No edit summary ย |
||
(10 intermediate revisions by 5 users not shown) | |||
Line 1: | Line 1: | ||
{{MitoPedia | {{MitoPedia | ||
|abbr= | |abbr=RP2-short | ||
|description=[[File:1OctM;2D;3P;4S;5U;6Rot-.png|300px]] | |description=[[File:1OctM;2D;3P;4S;5U;6Rot-.png|300px]] | ||
|info='''A: | |info='''A:''' when [[Malate-anaplerotic pathway control state |malate-anaplerotic activity]] is zero | ||
}} | }} | ||
{{MitoPedia concepts | |||
::: '''[[SUIT protocol pattern]]:''' | |mitopedia concept=MiP concept, SUIT protocol, Recommended | ||
}} | |||
{{MitoPedia methods | |||
|mitopedia method=Respirometry | |||
}} | |||
{{MitoPedia O2k and high-resolution respirometry}} | |||
{{MitoPedia topics}} | |||
::: '''[[SUIT protocol pattern]]:''' 1OctM;2D;3P;4S;5U;6Rot- | |||
The SUIT-005 protocols provide information on the [[Fatty_acid_oxidation_pathway_control_state|F-pathway]], the combined [[FN|FN]] pathway, and the convergence [[FNS |FNS]] pathways in the [[Oxidative phosphorylation|OXPHOS state]]. [[FNS |FNS]] comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum [[OXPHOS | The SUIT-005 protocols provide information on the [[Fatty_acid_oxidation_pathway_control_state|F-pathway]], the combined [[FN|FN]] pathway, and the convergence [[FNS |FNS]] pathways in the [[Oxidative phosphorylation|OXPHOS state]]. [[FNS |FNS]] comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum [[OXPHOS capacity| OXPHOS-]] and [[ET capacity |ET capacity]]. SUIT-005 can be extended with the CIV assay module. | ||
__TOC__ | __TOC__ | ||
ย Communicated by | ย Communicated by [[Gnaiger E]] (last update 2019-06-05) | ||
== Specific SUIT protocols == | == Specific SUIT protocols == | ||
Line 22: | Line 29: | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::+ Reasonable duration of the experiment (1.5 to 2 h); 2-3 h when the CIV module is added. | ::::+ Reasonable duration of the experiment (1.5 to 2 h); 2-3 h when the CIV module is added. | ||
:::+ The protocol provides information on F-OXPHOS capacity in the absence of a significant [[OXPHOS | ::::+ 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]]. | ||
:::+ [[FNS |FNS]] comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum [[OXPHOS | ::::+ [[FNS |FNS]] comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum [[OXPHOS capacity| OXPHOS-]] and [[ET capacity |ET capacity]]. | ||
:::+ PM & S reconstitutes TCA cycle activity ([[PMS]]) if the 2-oxoglutarate carrier does not outcompete the sources of 2-oxoglutarate. | ::::+ PM & S reconstitutes TCA cycle activity ([[PMS]]) if the 2-oxoglutarate carrier does not outcompete the sources of 2-oxoglutarate. | ||
:::+ Application of the [[cytochrome c]] test early in the protocol ensures comparability of all states in case of any effect of c | ::::+ Application of the [[cytochrome c]] test early in the protocol ensures comparability of all states in case of any effect of c. | ||
ย | |||
::::- 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]]). | |||
::::- SRot<sub>''E''</sub> may be underestimated if S is not saturating. | |||
::::- 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]]. | |||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
::::* [[SUIT-002]] check for malate-linked anaplerotic activity | ::::* [[SUIT-002]] or [[SUIT-025]] to measure F-OXPHOS capacity when malate anaplerotic activity is present; | ||
::::* [[SUIT-027]] to check for malate-linked anaplerotic activity | |||
::::* [[SUIT-017]] | |||
== References == | == References == | ||
Line 47: | Line 56: | ||
|sort=Was published in year | |sort=Was published in year | ||
|order=descending | |order=descending | ||
}} | }} |
Latest revision as of 14:30, 10 March 2023
Description
Abbreviation: RP2-short
Reference: A: when malate-anaplerotic activity is zero
MitoPedia concepts:
MiP concept,
SUIT protocol,
Recommended
MitoPedia methods:
Respirometry
- SUIT protocol pattern: 1OctM;2D;3P;4S;5U;6Rot-
The SUIT-005 protocols provide information on the F-pathway, the combined FN pathway, and the convergence FNS pathways in the OXPHOS state. FNS comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum OXPHOS- and ET capacity. SUIT-005 can be extended with the CIV assay module.
Communicated by Gnaiger E (last update 2019-06-05)
Specific SUIT protocols
SUIT-005 O2 pfi D11
- SUIT-005 O2 pfi D011 for permeabilized fibers
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 | F(N) | FAO | 1OCtM;2D;2c
|
3P | OctPMP | FN | F&CI | 1OctM;2D;2c;3P
|
4S | OctPMSP | FNS | F&CI&II | 1OctM;2D;2c;3P;4S
|
5U | OctPMSE | FNS | F&CI&II | 1OctM;2D;2c;3P;4S;5U
|
6Rot | SE | S | CII | 1OctM;2D;3P;4S;5U;6Rot
|
7Ama | ROX | 1OctM;2D;3P;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
- 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
- + Reasonable duration of the experiment (1.5 to 2 h); 2-3 h when the CIV module is added.
- + The protocol provides information on F-OXPHOS capacity in the absence of a significant OXPHOS capacity in the malate-anaplerotic pathway control state.
- + FNS comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum OXPHOS- and ET capacity.
- + PM & S reconstitutes TCA cycle activity (PMS) if the 2-oxoglutarate carrier does not outcompete the sources of 2-oxoglutarate.
- + Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c.
- - 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).
- - SRotE may be underestimated if S is not saturating.
- - 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.