Abbreviation: CCP-ce microalgae
Reference: A: Coupling control in intact cells without oligomycin; ce: non-permeabilized cells -SUIT-003
SUIT number: D039_ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D039 has been designed to study the coupling control state of intact microalgal cells. The different capacities are tested in the sequence OXPHOS, LEAK respiration and Electron transfer pathway. Depending on the species of microalgae studied, the oligomycin or any other inhibitor of the ATP synthase or of the adenine nucleotide translocase might not have an effect and the LEAK respiration cannot be achieved. Therefore, it is recommend to test the effect of these inhibitors in preliminary experiments for different species. Also, the inhibitory effect on the electron transfer system has to be studied to avoid an underestimation of the ET capacity. This protocol has been optimised specifically for the microalgae species Chlamydomonas reinhardtii in which oligomycin does not have an effect.
Communicated by Huete-Ortega M and Gnaiger E (last update 2021-03-23)
Steps and respiratory states
|Step||State||Pathway||Q-junction||Comment - Events (E) and Marks (M)|
|ce4Rot||B(Rot) or ROX||ce1;(ce2Omy);ce3U;ce4Rot
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- Pathway control
Strengths and limitations
- + Reasonable duration of the experiment.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
- - Depending on the species of microalgae studied oligomycin or any other inhibitor of the ATP synthase or of the adenine nucleotide translocase might not have an effect and therefore the LEAK respiration cannot be achieved.
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Chemicals and syringes
|Step||Chemical(s) and link(s)||Comments|
|(ce2Omy)||Oligomycin (Omy)||This step can be skipped|
|ce3U||Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U)||Can be substituted for other uncoupler|
|ce5Ama||Antimycin A (Ama)|
- Suggested stock concentrations are shown in the specific DL-Protocol.
MitoPedia methods: Respirometry