SUIT-006 O2 mt D022: Difference between revisions
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ย Communicated by [[Cardoso LH]], [[Komlodi T]], [[Huete-Ortega M]] and [[Doerrier C]] (last update 2019-03-05) ย | ย Communicated by [[Cardoso LH]], [[Komlodi T]], [[Huete-Ortega M]] and [[Doerrier C]] (last update 2019-03-05) ย | ||
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::::* [[SUIT-003]]: Coupling control protocol for non-permeabilized cells (ce), counterpart to SUIT-006 for mt-preparations (mtprep). | ::::* [[SUIT-003]]: Coupling control protocol for non-permeabilized cells (ce), counterpart to SUIT-006 for mt-preparations (mtprep). | ||
== References == | |||
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{{MitoPedia concepts | {{MitoPedia concepts |
Revision as of 18:38, 5 March 2019
Description
Abbreviation: S
Reference: A SUIT-006
SUIT number: D022_1SRot;2D;2c;3U;4Ama
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: S
- SUIT protocol pattern: linear coupling control protocol 1SRot;2D;2c;3U;4Ama
SUIT-006 O2 mt D022 is a coupling control protocol for for mitochondrial preparations (tissue homogenate and isolated mitochondria). Different coupling control states are assessed (L(n) - P - E) at succinate pathway control state.
Communicated by Cardoso LH, Komlodi T, Huete-Ortega M and Doerrier C (last update 2019-03-05)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1SRot | SL | S | CII | 1SRot
|
2D | SP | S | CII | 1SRot;2D
|
2c | ScP | S | CII | 1SRot;2D;2c
|
(3Omy) | ScL(Omy) | 1SRot;2D;2c;(3Omy)
| ||
4U | SE | S | CII | 1SRot;2D;2c;4U
|
5Ama | ROX | 1SRot;2D;2c;4U;5Ama
|
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- 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 is indicated when the aim is to analyse coupling control in mtprep (see MitoPedia: Sample preparations) at Succinate pathway control state.
- + The combination of succinate with rotenone, that inhibits complex I, provides the best coupling control protocol specific for S-pathway, avoiding reverse electron transfer (RET).
- + Reasonable duration of the experiment.
- + This protocol can be extended with the Complex IV module.
- - This protocol does not include internal ET-pathway control steps.
- - Careful washing is required after the experiment to avoid carry-over of uncoupler and inhibitors.
Compare SUIT protocols
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
Krumschnabel 2014 Methods Enzymol | 2014 | Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. https://doi.org/10.1016/B978-0-12-416618-9.00009-1 | Mouse | Nervous system |
MitoPedia concepts:
SUIT protocol,
SUIT A,
Find
MitoPedia methods:
Respirometry