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 ET. 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 2019-04-10)
Steps and respiratory states
|Step||State||Pathway||Q-junction||Comment - Events (E) and Marks (M)|
- » Selected keywords in SUIT protocols
- Pathway control
Strengths and limitations
- The SUIT-003 O2 ce D039 is a cell coupling control protocol. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce).
- CIV activity can also be measured after the cell viability test module.
- + 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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MitoPedia methods: Respirometry