SUIT-012 O2 ce-pce D052: Difference between revisions
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::: '''[[SUIT protocol pattern]]:''' diametral ce1;1Dig;1PM;2D;2c;3G;4U;5Ama | ::: '''[[SUIT protocol pattern]]:''' diametral ce1;1Dig;1PM;2D;2c;3G;4U;5Ama | ||
The SUIT-012 O2 pce D052 protocols specifically focus on assessing the linear coupling control ([[LEAK-respiration|''L'']]-[[Oxidative phosphorylation| ''P'']]-[[ET-capacity| ''E'']]) with NADH-linked substrates ([[PM pathway control state|PM]]) in permeabilized fibers. Addition of [[Glutamate|G]] in NADH-supported [[Oxidative phosphorylation| OXPHO]]S enables evaluating the [[glutamate anaplerotic pathway control state]]. SUIT-012 O2 pce D052 can be extended with the CIV assay module. | The SUIT-012 O2 ce-pce D052 protocols specifically focus on assessing the linear coupling control ([[LEAK-respiration|''L'']]-[[Oxidative phosphorylation| ''P'']]-[[ET-capacity| ''E'']]) with NADH-linked substrates ([[PM pathway control state|PM]]) in permeabilized fibers. Addition of [[Glutamate|G]] in NADH-supported [[Oxidative phosphorylation| OXPHO]]S enables evaluating the [[glutamate anaplerotic pathway control state]]. SUIT-012 O2 ce-pce D052 can be extended with the CIV assay module. | ||
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== References == | == References == | ||
{{#ask:[[Category:Publications]] [[Instrument and method::O2k-Protocol]] [[Additional label::SUIT-012 pce D052]] ย | {{#ask:[[Category:Publications]] [[Instrument and method::O2k-Protocol]] [[Additional label::SUIT-012 ce-pce D052]] ย | ||
|?Was published in year=Year | |?Was published in year=Year | ||
|?Has title=Reference | |?Has title=Reference |
Revision as of 14:52, 27 March 2019
Description
Abbreviation: N(PGM)
Reference: A SUIT-012
SUIT number: D052_ce1;1Dig;1PM;2D;2c;3G;4U;5Ama
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: N(PGM)
- SUIT protocol pattern: diametral ce1;1Dig;1PM;2D;2c;3G;4U;5Ama
The SUIT-012 O2 ce-pce D052 protocols specifically focus on assessing the linear coupling control (L- P- E) with NADH-linked substrates (PM) in permeabilized fibers. Addition of G in NADH-supported OXPHOS enables evaluating the glutamate anaplerotic pathway control state. SUIT-012 O2 ce-pce D052 can be extended with the CIV assay module.
Communicated by Huete-Ortega M, Iglesias-Gonzalez J, Komlodi T and Gnaiger E (last update 2019-03-27)
Steps and respiratory States
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
1Dig | REN | ce1;1Dig
|
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1PM | PML(n) | N | CI | 1PM
|
2D | PMP | N | CI | 1PM;2D
|
2c | PMcP | N | CI | 1PM;2D;2c
|
3G | PGMP | N | CI | 1PM;2D;2c;3G
|
4U | PGME | N | CI | 1PM;2D;2c;3G;4U
|
5Ama | ROX | 1PM;2D;2c;3G;4U;5Ama
|
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
- + This protocol allows to evaluate the function of the TCA cycle without the involvement of the complex II (S-pathway). Would be useful to understand the contribution and the activity of the dehydrogenases.
- + Mitochondrial external membrane integrity can be measured with the addition of cytochrome c. 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.
- + This protocol can be extended with the Complex IV module, which can prolong the experimental time with ~30 min.
- + GM and PM yield typically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM: the fraction of the N-pathway is lower and of the S-pathway is higher with GM compared to PM (GMP is inhibited by the CII inhibitor malonic acid to a larger extent than PMP). PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-pathway, since an impairment of N-pathway capacity can be compensated partially by activation of the S-pathway. This is an advantage compared to SUIT-011 for diagnosis of N-capacity.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
Compare SUIT protocols
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry