SUIT-003 O2 ce D062: Difference between revisions
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::: '''[[MitoPedia: SUIT]]''' - Coupling control protocol (CCP) in living cells (ce) with the carrier of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv): DMSO. | ::: '''[[MitoPedia: SUIT]]''' - Coupling control protocol (CCP) in living cells (ce) with the carrier of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv): DMSO. | ||
::: '''[[SUIT protocol pattern]]:''' ce1;ce1DMSO;(ce2Omy);ce3U;ce4Rot;ce5Ama | ::: '''[[SUIT protocol pattern]]:''' ce1;ce1DMSO;(ce2Omy);ce3U;ce4Rot;ce5Ama | ||
SUIT-003 O2 ce D062 protocol is the control of the [[SUIT-003 O2 ce D050]] adding the carrier DMSO in [[ROUTINE]] respiration instead of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv). Moreover,the SUIT-003 O2 ce D062 protocol allows to evaluate the [[coupling control state |coupling control]] of living cells. Respiratory capacities are tested in a sequence of coupling states: [[ROUTINE]] and [[ET]]. | SUIT-003 O2 ce D062 protocol is the control of the [[SUIT-003 O2 ce D050]] adding the carrier DMSO in [[ROUTINE]] respiration instead of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv). Moreover,the SUIT-003 O2 ce D062 protocol allows to evaluate the [[coupling control state |coupling control]] of living cells. Respiratory capacities are tested in a sequence of coupling states: [[ROUTINE]] and [[ET]]. Optionally, to study [[LEAK-respiration]], the [[phosphorylation system]] is inhibited by [[oligomycin]]. The final concentration of [[oligomycin]] has to be carefully optimized for various cell types, to minimize the inhibitory effect on the [[Electron transfer-pathway|electron transfer system]] which would lead to an underestimation of [[ET-capacity]]. ย | ||
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Revision as of 16:04, 25 June 2019
Description
Abbreviation: Carrier for MitoKit-CII/Plasma membrane-permeable succinate (NV118)
Reference: A: - SUIT-003
SUIT number: D062_ce1;ce1DMSO;(ce2Omy);ce3U;ce4Rot;ce5Ama
O2k-Application: O2
- MitoPedia: SUIT - Coupling control protocol (CCP) in living cells (ce) with the carrier of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv): DMSO.
- SUIT protocol pattern: ce1;ce1DMSO;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D062 protocol is the control of the SUIT-003 O2 ce D050 adding the carrier DMSO in ROUTINE respiration instead of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv). Moreover,the SUIT-003 O2 ce D062 protocol allows to evaluate the coupling control of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE and ET. Optionally, to study LEAK-respiration, the phosphorylation system is inhibited by oligomycin. The final concentration of oligomycin has to be carefully optimized for various cell types, to minimize the inhibitory effect on the electron transfer system which would lead to an underestimation of ET-capacity.
Communicated by Doerrier C (last update 2019-06-25)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M)
|
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce1DMSO | ROUTINE | ce1
| ||
(ce2Omy) | L(Omy) | ce1;ce1Snv;(ce2Omy)
| ||
ce3U | E | ce1;ce1Snv;(ce2Omy);ce3U
| ||
ce4Rot | ROX | ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot
| ||
ce5Ama | ROX | ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot;ce5Ama
|
- 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
- + Reasonable duration of the experiment.
- - To obtain LEAK respiration, the concentration of oligomycin has to be carefully tested to avoid inhibitory effects in the evaluation of the ET-capacity.
Compare SUIT protocols
- SUIT-003 O2 ce D050 the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv) is added in ROUTINE.
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Respirometry