SUIT-003 O2 ce D039: Difference between revisions
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|SUIT number=D039_ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama | |SUIT number=D039_ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama | ||
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SUIT | {{Template:SUIT text D039}} | ||
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Communicated by [[Huete-Ortega M]] and [[Gnaiger E]] (last update 2021-03-23) | |||
== Representative traces == | |||
[[File:D009 Traces algae.png|600px]] | |||
{{Template:SUIT-003 O2 ce D039}} | {{Template:SUIT-003 O2 ce D039}} | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::* The [[SUIT-003 O2 ce D039]] is a cell [[coupling control protocol]]. The total cell count (''N''<sub>ce</sub>) is the sum of viable cells (''N''<sub>vce</sub>) and dead cells (''N''<sub>dce</sub>). | :::* The [[SUIT-003 O2 ce D039]] is a cell [[coupling-control protocol]]. The total cell count (''N''<sub>ce</sub>) is the sum of viable cells (''N''<sub>vce</sub>) and dead cells (''N''<sub>dce</sub>). | ||
:::* CIV activity can also be measured after the cell viability test module. | :::* CIV activity can also be measured after the cell viability test module. | ||
:::+ Reasonable duration of the experiment. | :::+ Reasonable duration of the experiment. | ||
:::- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler. | :::- 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 | :::- 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. | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
== Chemicals and syringes == | |||
{{Template:Chemicals SUIT-003 D039}} | |||
::: Suggested stock concentrations are shown in the specific DL-Protocol. | |||
== References == | == References == |
Latest revision as of 14:47, 23 March 2021
Description
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
O2k-Application: O2
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)
Representative traces
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
(ce2Omy) | L(Omy) | ce1;(ce2Omy)
| ||
ce3U | E | ce1;(ce2Omy);ce3U
| ||
ce4Rot | B(Rot) or ROX | ce1;(ce2Omy);ce3U;ce4Rot
| ||
ce5Ama | ROX | ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- ยป Electron transfer pathway
- ยป Fatty acid oxidation pathway control state, F
- ยป NADH electron transfer-pathway state, N
- ยป Succinate pathway control state, S
- ยป NS-pathway control state, NS
- ยป Glycerophosphate pathway control state, Gp
- ยป Complex IV single step, CIV
- ยป Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- ยป Glutamate, G
- ยป Glycerophosphate, Gp
- ยป Malate, M
- ยป Octanoylcarnitine, Oct
- ยป Pyruvate, P
- ยป Succinate, S
- Main fuel substrates
- Glossary
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.
Compare SUIT protocols
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 |
ce4Rot | Rotenone (Rot) | |
ce5Ama | Antimycin A (Ama) |
- Suggested stock concentrations are shown in the specific DL-Protocol.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry