From Bioblast
Description
Abbreviation: AOX-ce SHAM+CN
Reference: A: Determination of respiration through the CIII-CIV pathway
- SUIT protocol pattern: ce1;ce2SHAM;ce3KCN
The SUIT-023 provides an estimation of mitochondrial respiration in living cells through the CIII- cytochrome c oxidase pathway (CIII-CIV), which is inhibited by antimycin A and cyanide. This protocol must be complemented with the study in parallel of mitochondrial respiration through the Q-alternative oxidase pathway (Q-AOX; see SUIT-022).
Communicated by Huete-Ortega M, Gnaiger E (last update 2019-06-05)
Specific SUIT protocols
SUIT-023 O2 ce D053
- SUIT-023 O2 ce D053 for living microalgal cells (ce)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce2SHAM | ROUTINE OR ROX | ce1;ce2SHAM
| ||
ce3KCN | ROX | ce1;ce2SHAM;ce3KCN
|
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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
- SUIT-023 provides an estimation of the cyanide-and antimycin A-non-resistant mitochondrial respiration due to the activity cytochrome c oxidase (CIV) in cells. To complement the study with the estimation of the mitochondrial respiration through the Q-alternative oxidase pathway, it must be carried out in parallel to SUIT-022.
- + This protocol can be used with many types of cells, although optimal concentrations of inhibitors have to be tested.
- + Short duration of experiment
- - No estimation of coupling control
- - The use of cyanide requires careful cleaning of the chambers after use and careful handling of the chemical to guarantee health safety.
Compare SUIT protocols
- SUIT-022 O2 ce D051 for microalgal living cells
References
MitoPedia concepts: SUIT protocol, Recommended
MitoPedia methods:
Respirometry