SUIT-003 O2 ce D038: Difference between revisions
No edit summary |
No edit summary |
||
Line 4: | Line 4: | ||
|info='''A''' Coupling control protocol and Crabtree effect test -'''[[SUIT-003]]''' | |info='''A''' Coupling control protocol and Crabtree effect test -'''[[SUIT-003]]''' | ||
|application=O2 | |application=O2 | ||
|SUIT number=D038_ce1; | |SUIT number=D038_ce1;Omy;ce3U;ce3Glc;ce3'U;ce4Ama | ||
}} | }} | ||
::: '''[[MitoPedia: SUIT]]''' | ::: '''[[MitoPedia: SUIT]]''' | ||
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' ceGlc | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' ceGlc | ||
::: '''[[SUIT protocol pattern]]:''' complex cell [[coupling control protocol]] ce1; | ::: '''[[SUIT protocol pattern]]:''' complex cell [[coupling control protocol]] ce1;Omy;ce3U;ce3Glc;ce3'U- | ||
SUIT-003 O2 ce D038 has been designed to study the [[coupling control state]] and respiration | SUIT-003 O2 ce D038 has been designed to study the regulation of [[coupling control state]] and respiration by glucose (Crabtree effect) of intact cells on [[ET-capacity]]. To obtain more information about Crabtree effect, this protocol can be used in parallel with [[SUIT-003 O2 ce D037]]. SUIT-003 O2 ce D038 follows a modular approach and can be easily used in combination with the assessment of complex IV capacity and respirometric cell viability test. This protocol allows us to omit oligomycin, therefore the LEAK-respiration and measure the ET-capacity directly after [[ROUTINE]]. | ||
__TOC__ | __TOC__ | ||
ย Communicated by [[Garcia-Souza LF]], [[Cardoso LH]] and [[Gnaiger E]] (last update 2019-04- | ย Communicated by [[Komlodi T ]], [[Garcia-Souza LF]], [[Cardoso LH]] and [[Gnaiger E]] (last update 2019-04-03) | ||
{{Template:SUIT-003 O2 ce D038}} | {{Template:SUIT-003 O2 ce D038}} | ||
Line 23: | Line 23: | ||
:::* 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 D037]]. In SUIT-003 O2 ce D037, the addition of glucose during ROUTINE state can lead to increase in ATP production by glycolysis, which can indirectly decrease respiration. In SUIT-003 O2 ce D038, with addition of glucose in ET state, this effect is bypassed, and only the direct effect of glucose on the electron | :::+ To obtain more information about Crabtree effect, this protocol can be used in parallel with [[SUIT-003 O2 ce D037]]. In SUIT-003 O2 ce D037, the addition of glucose during ROUTINE state can lead to increase in ATP production by glycolysis, which can indirectly decrease respiration. In SUIT-003 O2 ce D038, with 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 inhibitors and uncoupler. | :::- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler. | ||
Revision as of 09:48, 3 April 2019
Description
Abbreviation: ceGlc
Reference: A Coupling control protocol and Crabtree effect test -SUIT-003
SUIT number: D038_ce1;Omy;ce3U;ce3Glc;ce3'U;ce4Ama
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: ceGlc
- SUIT protocol pattern: complex cell coupling control protocol ce1;Omy;ce3U;ce3Glc;ce3'U-
SUIT-003 O2 ce D038 has been designed to study the regulation of coupling control state and respiration by glucose (Crabtree effect) of intact cells on ET-capacity. To obtain more information about Crabtree effect, this protocol can be used in parallel with SUIT-003 O2 ce D037. SUIT-003 O2 ce D038 follows a modular approach and can be easily used in combination with the assessment of complex IV capacity and respirometric cell viability test. This protocol allows us to omit oligomycin, therefore the LEAK-respiration and measure the ET-capacity directly after ROUTINE.
Communicated by Komlodi T , Garcia-Souza LF, Cardoso LH and Gnaiger E (last update 2019-04-03)
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
| ||
ce3Glc | E | ce1;(ce2Omy);ce3U;ce3Glc
| ||
ce3'U | E | ce1;(ce2Omy);ce3U;ce3Glc;ce3'U
| ||
ce4Ama | ROX | ce1;(ce2Omy);ce3U;ce3Glc;ce3'U;ce4Ama
|
- 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 D038 is a cell coupling control protocol. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce).
- This protocol can be extended with the cell viability test module (See CCV SUIT-003 O2 ce-pce D020).
- 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 D037. In SUIT-003 O2 ce D037, the addition of glucose during ROUTINE state can lead to increase in ATP production by glycolysis, which can indirectly decrease respiration. In SUIT-003 O2 ce D038, with 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 inhibitors and uncoupler.
Compare SUIT protocols
- SUIT-003 O2 ce D037 can be used in parallel to obtain more information on Crabtree effect.
- SUIT-003 O2 ce-pce D013 for assessment of coupling control, Crabtree effect and cell viability test.
- SUIT-003 O2 ce-pce D018 for assessment of coupling control, Crabtree effect, malate effect and cell viability test.
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry