SUIT-012: Difference between revisions
From Bioblast
No edit summary |
No edit summary |
||
Line 11: | Line 11: | ||
[[File:1PM;2D;2c;3G;4U;5Ama.png|300px]] ย | [[File:1PM;2D;2c;3G;4U;5Ama.png|300px]] ย | ||
* [[SUIT-012 O2 mt D027]] for isolated mitochondria | * [[SUIT-012 O2 mt D027]] for isolated mitochondria | ||
[[File:ce1;1Dig;1PM;2D;2c;3G;4U;5Ama.png| | [[File:ce1;1Dig;1PM;2D;2c;3G;4U;5Ama.png|400px]] | ||
* [[SUIT-012 O2 pce D###]] for permeabilized cells | * [[SUIT-012 O2 pce D###]] for permeabilized cells | ||
== References == | == References == |
Revision as of 16:26, 28 January 2019
Description
Abbreviation: N(PGM)
Reference: A ยปVersions
- SUIT-category: N(PGM)
- SUIT protocol pattern: diametral 1PM;2D;2c;3G;4U-
Communicated by Cardoso LH, Doerrier C, Huete-Ortega M, Iglesias-Gonzalez J and Gnaiger E (last update 2019-01-28)
DLP applications
- SUIT-012 O2 mt D027 for isolated mitochondria
- SUIT-012 O2 pce D### for permeabilized cells
References
Steps and respiratory states
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 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.
- + Complex IV activity can be measured.
- + 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
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry