Difference between revisions of "SUIT-003 pH ce D067"
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{{MitoPedia | {{MitoPedia | ||
|abbr=CCP-Crabtree | |abbr=CCP-Crabtree with glycolysis inhibition | ||
|description=[[.png|350px]] | |description=[[File:D067 diagram.png|350px]] | ||
|info='''A''' [[Coupling control protocol]] and [[Crabtree effect]] - '''[[SUIT-003]]''' | |info='''A''' [[Coupling control protocol]] and [[Crabtree effect]] with glycolysis inhibition- '''[[SUIT-003]]''' | ||
|application=O2 | |application=O2 | ||
|SUIT number=D067_ce1;ce1Glc;ce2Omy;ce3DG | |SUIT number=D067_ce1;ce1Glc;ce2Omy;ce3DG;ce4Ama | ||
}} | }} | ||
::: '''[[MitoPedia: SUIT]]''' | ::: '''[[MitoPedia: SUIT]]''' | ||
::: '''[[SUIT protocol pattern]]:''' ce1;ce1Glc;ce2Omy;ce3DG- | ::: '''[[SUIT protocol pattern]]:''' ce1;ce1Glc;ce2Omy;ce3DG;- | ||
{{Template:SUIT text D067}} | {{Template:SUIT text D067}} | ||
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== Representative traces == | == Representative traces == | ||
[[.png|600px]] | [[File:D067 O2 trace.png|600px]][[File:D067 pH trace.png|600px]] | ||
{{Template:SUIT-003 O2 ce D067}} | {{Template:SUIT-003 O2 ce D067}} | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::* The SUIT-003 O2 ce | :::* The SUIT-003 O2 ce D067 is protocol for living cells to study Crabtree effect and glycolysis inhibition by measuring respiration and proton flux simultaneously. | ||
:::* 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. | ||
:::+ To obtain more information about Crabtree effect, this protocol can be used in parallel with [[SUIT-003 O2 ce D038]] | :::+ To obtain more information about Crabtree effect, this protocol can be used in parallel with [[SUIT-003 O2 ce D038]] and [[SUIT-003 O2 ce D037]]. Addition of glucose in the ROUTINE state can lead to an increase in ATP production by glycolysis, which can indirectly decrease respiration. In SUIT-003 O2 ce D038, with the addition of glucose in ET state, this effect is bypassed, and only the direct effect of glucose on the [[Electron transfer-pathway|electron transfer system]] is tested. | ||
:::- Careful washing is required after the experiment to avoid carry-over of | :::- Careful washing is required after the experiment to avoid carry-over of Omy. | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
::::* [[SUIT-003 O2 ce D038]] and [[SUIT-003 O2 ce D037]] | ::::* [[SUIT-003 O2 ce D038]] and [[SUIT-003 O2 ce D037]] for assessment for assessment of coupling control and Crabtree effect. | ||
::::* [[SUIT-003 O2 ce-pce D013]] for assessment of coupling control, Crabtree effect, malate effect, and cell viability test. | ::::* [[SUIT-003 O2 ce-pce D013]] for assessment of coupling control, Crabtree effect, malate effect, and cell viability test. | ||
::::* [[SUIT-003 O2 ce-pce D018]] for assessment of coupling control, Crabtree effect, and cell viability test. | ::::* [[SUIT-003 O2 ce-pce D018]] for assessment of coupling control, Crabtree effect, and cell viability test. |
Revision as of 10:56, 21 February 2020
Description
Abbreviation: CCP-Crabtree with glycolysis inhibition
Reference: A Coupling control protocol and Crabtree effect with glycolysis inhibition- SUIT-003
SUIT number: D067_ce1;ce1Glc;ce2Omy;ce3DG;ce4Ama
O2k-Application: O2
- MitoPedia: SUIT
- SUIT protocol pattern: ce1;ce1Glc;ce2Omy;ce3DG;-
The SUIT-003 O2 ce D067 is designed to study the effect of glucose (or fructose; Crabtree effect) on ROUTINE respiration and glycolysis inhibition on LEAK-respiration simultaneously measured with proton flux in living cells. If the aim is to analyze only the Crabtree effect and the effect of 2-Deoxyglucose on proton flux and respiration via glycolysis inhibition, Oligomycin can be omitted. If there is an interest in studying coupling control, oligomycin should be added to promote LEAK-respiration. To obtain more information about the Crabtree effect, it is ideal to use in parallel the protocol SUIT-003 O2 ce D038 (glucose addition in ET state) and SUIT-003 O2 ce D037 (glucose addition in ROUTINE). The latter can be extended by the cell viability test and the Complex IV module.
Communicated by Komlodi T, [ Gnaiger E (last update 2020-02-20)
Representative traces
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce1Glc | ROUTINE | ce1;ce1Glc
| ||
ce2Omy | LOmy | ce1;ce1Glc;ce2Omy
| ||
ce3DG | L | ce1;ce1Glc;ce2Omy;ce3DG
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- The SUIT-003 O2 ce D067 is protocol for living cells to study Crabtree effect and glycolysis inhibition by measuring respiration and proton flux simultaneously.
- CIV activity can also be measured after the cell viability test module.
- + Reasonable duration of the experiment.
- + To obtain more information about Crabtree effect, this protocol can be used in parallel with SUIT-003 O2 ce D038 and SUIT-003 O2 ce D037. Addition of glucose in the ROUTINE state can lead to an increase in ATP production by glycolysis, which can indirectly decrease respiration. In SUIT-003 O2 ce D038, with the addition of glucose in ET state, this effect is bypassed, and only the direct effect of glucose on the electron transfer system is tested.
- - Careful washing is required after the experiment to avoid carry-over of Omy.
Compare SUIT protocols
- SUIT-003 O2 ce D038 and SUIT-003 O2 ce D037 for assessment for assessment of coupling control and Crabtree effect.
- SUIT-003 O2 ce-pce D013 for assessment of coupling control, Crabtree effect, malate effect, and cell viability test.
- SUIT-003 O2 ce-pce D018 for assessment of coupling control, Crabtree effect, and cell viability test.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry