Difference between revisions of "SUIT-005 O2 pfi D011"
From Bioblast
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== Strengths and limitations == | == Strengths and limitations == | ||
:::+ The protocol provides information on FAO capacity in the absence of other, potentially interfering pathways, both in the LEAK state and in OXPHOS. | |||
:::+ FNS OXPHOS capacity comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity. | |||
:::+ FNS ET capacity is a good estimate of overall ET capacity in many cell types. | |||
:::+ The presence of PM and S establishes a fully operative 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. | |||
:::+ Reasonable duration of the experiment. | |||
:::- F OXPHOS capacity may be underestimated. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) + malate increased OXPHOS by 26% (Lemuieux et al 2011). | |||
:::- SRot(E) may be underestimated if S is not saturating. | |||
:::- CIV activity is not measured, to save experimental time. | |||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
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::::* [[SUIT-002]] | ::::* [[SUIT-002]] | ||
::::* [[SUIT-002 O2 pfi D006]] | ::::* [[SUIT-002 O2 pfi D006]] | ||
::::* [[Isolated mitochondria |imt]], ROX: 1OctM;2D;2c;3P;4S;5U;6Rot;7Ama | |||
::::* [[Permeabilized muscle fibres |pfi]], high O<sub>2</sub>, no ROX: FNS(PM)01_pfiO2,1OctM,2D(c),3P,4S,5U,6Rot | |||
Revision as of 11:53, 23 January 2019
Description
Abbreviation: FNS(Oct,PM)
Reference: A: SUIT-005 short protocol linked to SUIT-002 O2 pfi D006 - SUIT RP2 (human skeletal muscle)
SUIT number: D011_1OctM;2D;3P;4S;5U;6Rot
O2k-Application: O2
- SUIT-category: FNS(Oct,PM)
- SUIT protocol pattern: diametral 1OctM;2D;3P;4S;5U;6Rot
- Linked to SUIT-002 O2 pfi D006 - SUIT RP2, specifically for human skeletal muscle mitochondria.
- Harmonized with SUIT-004 O2 pfi D010.
- F-pathway has to be tested previously.
Communicated by Doerrier C, Iglesias-Gonzalez J and Gnaiger E (last update 2019-01-21)
References
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
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- + The protocol provides information on FAO capacity in the absence of other, potentially interfering pathways, both in the LEAK state and in OXPHOS.
- + FNS OXPHOS capacity comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
- + FNS ET capacity is a good estimate of overall ET capacity in many cell types.
- + The presence of PM and S establishes a fully operative 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.
- + Reasonable duration of the experiment.
- - F OXPHOS capacity may be underestimated. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) + malate increased OXPHOS by 26% (Lemuieux et al 2011).
- - SRot(E) may be underestimated if S is not saturating.
- - CIV activity is not measured, to save experimental time.
Compare SUIT protocols
- SUIT-002
- SUIT-002 O2 pfi D006
- imt, ROX: 1OctM;2D;2c;3P;4S;5U;6Rot;7Ama
- pfi, high O2, no ROX: FNS(PM)01_pfiO2,1OctM,2D(c),3P,4S,5U,6Rot
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry