Description
Abbreviation: Plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv)
Reference: A: - SUIT-003
SUIT number: D050_ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot;ce5Ama
O2k-Application: O2
- MitoPedia: SUIT - Plasma membrane-permeable succinate (NV118) in ROUTINE respiration. Coupling control protocol (CCP) in living cells (ce)
- SUIT protocol pattern: ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D050 protocol is designed to study CI-deficiency in living cells. Plasma membrane-permeable succinate (Snv), which is added in ROUTINE stimulates mitochondrial respiration in CI-deficient human blood cells, fibroblasts and heart fibres, acting on Complex II of the electron transfer system. Moreover, the SUIT-003 O2 ce D050 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
| ||
ce1Snv | 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
|
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- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» 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 D062 carrier control SUIT protocol for SUIT-003 O2 ce D050 in living cells. The carrier of the plasma membrane-permeable succinate (DMSO) is added in ROUTINE.
- SUIT-003 O2 ce D060 CCP with cell permeable compounds (MitoKit-CII/Succinate-nv and MitoKit-CII/Malonate-nv).
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry