From Bioblast
Description
Abbreviation: AOX-ce CN+SHAM
Reference: A Determination of the respiration due to the alternative oxidase pathway -SUIT-022
SUIT number: D051_ce1;ce2KCN;ce3SHAM
O2k-Application: O2
The SUIT-022 O2 ce D051 provides an estimation of mitochondrial respiration due to the Q-alternative oxidase pathway (Q-AOX pathway), which is cyanide-and antimycin A-resistant, for the specific case of microalgal cells. This protocol must be complemented with the study in parallel of mitochondrial respiration through the CIII-cytochrome c oxidase pathway (CIII-CIV; see SUIT-023).
Communicated by Huete-Ortega M and Gnaiger E (last update 2024-09-19)
Representative traces
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 O2 ce D051 provides an estimation of the cyanide-and antimycin A-resistant mitochondrial respiration due to the activity of the alternative oxidase in microalgal cells.
- The concentrations of inhibitors used in this protocol have been optimized for the specific case of the microalgal model species Chlamydomonas reinhardtii.
- + Short duration of experiment.
- - No estimation of coupling control.
- - The use of cyanide requires a 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 has to be used in parallel to obtain complete information of the contribution of both Q-AOX and CIII-CIV pathways to mitochondrial respiration.
Chemicals and syringes
Step | Chemical(s) and link(s) | Comments |
---|---|---|
ce2KCN | Cyanide (KCN) | |
ce3SHAM | Salicylhydroxamic acid (SHAM) |
|
- Suggested stock concentrations are shown in the specific DL-Protocol.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry