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MitoPedia | Terms and abbreviations | Preprints and history | MiP and biochemistry | Concepts and methods | MitoPedia: SUIT | MitoPedia: O2k |
SUIT-015
Description
Abbreviation: F+G+P+S_OXPHOS+Rot_ET
Reference: A: F-pathway in LEAK state and OXPHOS state
- SUIT protocol pattern: 1OctM;2D;3G;4P;5S;6U:7Rot-
SUIT-015 gives information on F-pathway in LEAK state and OXPHOS state avoiding FAO overestimation in the presence of anaplerotic pathways. In addition, the pathway control of FN and FNS in OXPHOS state and of FNS and S in ET state can be evaluated. SUIT-015 can be extended with the CIV assay module.
Communicated by Doerrier C, Huete-Ortega M, Gnaiger E (last update 2019-06-05)
Specific SUIT protocols
- SUIT-015 O2 pti D043 for permeabilized tissue
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1OctM | OctML | F(N) | CETF | 1OctM
|
2D | OctMP | F(N) | CETF | 1OctM;2D
|
3G | OctGMP | FN | CETF&CI | 1OctM;2D;3G
|
3c | OctGMcP | FN | CETF&CI | 1OctM;2D;3G;3c
|
4P | OctPGMP | FN | CETF&CI | 1OctM;2D;3G;3c;4P
|
5S | OctPGMSP | FNS | CETF&CI&II | 1OctM;2D;3G;3c;4P;5S
|
6U | OctPGMSE | FNS | CETF&CI&II | 1OctM;2D;3G;3c;4P;5S;6U
|
7Rot | SE | S | CII | 1OctM;2D;3G;3c;4P;5S;6U;7Rot
|
8Ama | ROX | 1OctM;2D;3G;3c;4P;5S;6U;7Rot;8Ama
|
Step | Respiratory state | Pathway control | ET-Complex entry into Q-junction | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | ROX |
- 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 PMG and S establishes a fully operative TCA cycle activity.
- + Reasonable duration of the experiment.
- - Application of the cytochrome c test after 3G in the protocol does not ensure comparability of all states in case of any effect of c. However, it allows to analyse whether the outer mt-membrane integrity is compromised.
- - F OXPHOS capacity may be underestimated. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) + malate increased OXPHOS by 26% (Lemieux 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
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
Schoepf 2016 FEBS J | 2016 | Schöpf B, Schäfer G, Weber A, Talasz H, Eder IE, Klocker H, Gnaiger E (2016) Oxidative phosphorylation and mitochondrial function differ between human prostate tissue and cultured cells. https://doi.org/10.1111/febs.13733 | Human | Endothelial;epithelial;mesothelial cell Genital Other cell lines Fibroblast |
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry