SUIT-006 O2 mt D022: Difference between revisions
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|abbr=S | |abbr=S | ||
|description=[[File:1SRot;2D;2c;3U;4Ama.png|400px]] | |description=[[File:1SRot;2D;2c;3U;4Ama.png|400px]] | ||
|info='''A''' '''[[SUIT-006]]''' | |info='''A''': Protocol for coupling control assessment in mitochondrial preparations (isolated mitochondria, tissue homogenate and permeabilized cells) '''[[SUIT-006]]''' | ||
|application=O2 | |application=O2 | ||
|SUIT number=D022_1SRot;2D;2c;3U;4Ama | |SUIT number=D022_1SRot;2D;2c;3U;4Ama | ||
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::: '''[[SUIT protocol pattern]]:''' linear [[coupling control protocol]] 1SRot;2D;2c;3U;4Ama | ::: '''[[SUIT protocol pattern]]:''' linear [[coupling control protocol]] 1SRot;2D;2c;3U;4Ama | ||
SUIT-006 O2 mt D022 is a [[coupling control protocol]] | SUIT-006 O2 mt D022 is a [[coupling control protocol]] for [[mitochondrial preparations]] (tissue homogenate, isolated mitochondria and permeabilized cells). Different coupling control states are assessed ([[LEAK-respiration|''L''(n)]] - [[Oxidative phosphorylation| ''P'']] - [[ET-capacity| ''E'']]) at [[Succinate pathway control state|succinate pathway control state]]. The use of [[rotenone]], that inhibits complex I, provides a specific coupling control protocol for S-pathway, avoiding [[Reverse electron flow from CII to CI|reverse electron transfer (RET)]]. | ||
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== Strengths and limitations == | == Strengths and limitations == | ||
:::* This protocol is | :::* This protocol is recommended when the aim is to analyse coupling control in [[mitochondrial preparations |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). | :::+ 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. | :::+ Reasonable duration of the experiment. |
Revision as of 11:34, 27 March 2019
Description
Abbreviation: S
Reference: A: Protocol for coupling control assessment in mitochondrial preparations (isolated mitochondria, tissue homogenate and permeabilized cells) 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 mitochondrial preparations (tissue homogenate, isolated mitochondria and permeabilized cells). Different coupling control states are assessed (L(n) - P - E) at succinate pathway control state. The use of rotenone, that inhibits complex I, provides a specific coupling control protocol for S-pathway, avoiding reverse electron transfer (RET).
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 recommended 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,
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MitoPedia methods:
Respirometry