From Bioblast
Description
Abbreviation: AOX (ce CN+SHAM)
Reference: A: Determination of the respiration due to the alternative oxidase pathway
- SUIT protocol pattern: ce1;ce2KCN;ce3SHAM
The SUIT-022 provides an estimation of mitochondrial respiration in cells through the Q-alternative oxidase pathway (Q-AOX), which is antimycin A- and cyanide resistant. This protocol must be complemented with the study in parallel of mitochondrial respiration due to the CIII- cytochrome c oxidase pathway (CIII-CIV; see SUIT-023).
Communicated by Huete-Ortega M, Gnaiger E (last update 2019-06-05)
Specific SUIT protocols
SUIT-022 O2 ce D051
- SUIT-022 O2 ce D051 for living microalgal cells (ce)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce2KCN | ROUTINE or ROX | ce1;ce2KCN
| ||
ce3SHAM | ROX | ce1;ce2KCN;ce3SHAM
|
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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-022 provides an estimation of the cyanide-and antimycin A-resistant mitochondrial respiration due to the activity of alternative oxidase in cells on ROUTINE. To complement the study with the estimation of mitochondrial respiration due to the CIII-CIV pathway, it must be carried out in parallel to SUIT-023.
- + This protocol can be used with many types of cells, although optimal concentrations of inhibitors have to be determined.
- + 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-023 O2 ce D053 for microalgal intact cells
References
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry