SUIT-006 O2 ce-pce D029: Difference between revisions
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|SUIT number=D029_ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama | |SUIT number=D029_ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama | ||
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::: [[MitoPedia: SUIT]] ย | ::: '''[[MitoPedia: SUIT]]''' | ||
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' N(PM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' N(PM) | ||
::: '''[[SUIT protocol pattern]]:''' Bidirectional linear [[coupling control protocol]] ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama | ::: '''[[SUIT protocol pattern]]:''' Bidirectional linear [[coupling control protocol]] ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama |
Revision as of 13:21, 6 March 2019
Description
Abbreviation: N(PM)
Reference: A to analyse the coupling control on permeabilized cells in N(PM) pathway control state- SUIT-006
SUIT number: D029_ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: N(PM)
- SUIT protocol pattern: Bidirectional linear coupling control protocol ce1;1Dig;1PM;2D;2c;3Omy;4U;5Ama
SUIT-006 O2 pce D029 is a coupling control protocol for permeabilized cells. Different coupling control states are assessed (L(n) - P - E or L(n) - P - L(Omy) - E) at N(PM) pathway control state. The use of oligomycin is optional, however, it provides important information when ATPases are present, a situation that may lead to overestimated L(n) respiration.
Communicated by Cardoso LH, Komlodi T and Doerrier C (last update 2019-03-05)
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
|
3Omy | PMLOmy | N | CI | 1PM;2D;2c;3Omy
|
4U | PME | N | CI | 1PM;2D;2c;3Omy;4U
|
5Ama | ROX | 1PM;2D;2c;3Omy;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 permeabilized cells in NADH-pathway using PM as substrates.
- + Reasonable duration of the experiment.
- + This protocol can be extended with the Complex IV module.
- - This protocol analyses the coupling control in NADH-pathway (with PM), however, it is less specific than a similar protocol containing the complex II inhibitor malonic acid, that would allow to analyse the NADH-pathway without any contribution of the S-pathway.
- - 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, if used.
Compare SUIT protocols
- SUIT-001 O2 pce D003: Extended protocol with SUIT-006 as the initial coupling control module, for permeabilized cells.
- SUIT-003: Coupling control protocol for non-permeabilized cells (ce), counterpart to SUIT-006 for mt-preparations (mtprep).
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry