SUIT-026 AmR mt D064: Difference between revisions
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|abbr= | |abbr=RET | ||
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|SUIT number=D064_1S;2Rot;3D;4Ama | |SUIT number=D064_1S;2Rot;3D;4Ama | ||
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::: '''[[MitoPedia: SUIT]]''' - Protocol for Evaluation of ROS production by RET in isolated mitochondria | |||
::: '''[[SUIT protocol pattern]]:'''ย 1S;2Rot;3D;4Ama | ::: '''[[SUIT protocol pattern]]:'''ย 1S;2Rot;3D;4Ama | ||
SUIT-026 AmR mt D064 has been designed to study the [[Reverse_electron_flow_from_CII_to_CI| RET]] production of ROS in isolated mitochondria. The protocol is focused on [[LEAK]] state for the [[S-pathway]] to provide the conditions of high membrane potential, redox power in the Q-junction and no presence of reduced NADH. Under these conditions, in several samples has been described that a reverse flow of the electrons into the membrane arm and the quinone binding site of the Complex I occurs, promoting the production of ROS. The addition of [[rotenone]] provides excellent control to this mechanism because this compound blocks the point on which the electrons leak from the Complex I. The last step, the titration of ADP at saturating concentrations to reach [[OXPHOS]] allow us to harmonize this protocol with [[SUIT-026 O2 mt D063]] for quality control. | |||
Revision as of 12:33, 26 June 2019
Description
Abbreviation: RET
Reference: A - SUIT-026
SUIT number: D064_1S;2Rot;3D;4Ama
O2k-Application: AmR
- MitoPedia: SUIT - Protocol for Evaluation of ROS production by RET in isolated mitochondria
- SUIT protocol pattern: 1S;2Rot;3D;4Ama
SUIT-026 AmR mt D064 has been designed to study the RET production of ROS in isolated mitochondria. The protocol is focused on LEAK state for the S-pathway to provide the conditions of high membrane potential, redox power in the Q-junction and no presence of reduced NADH. Under these conditions, in several samples has been described that a reverse flow of the electrons into the membrane arm and the quinone binding site of the Complex I occurs, promoting the production of ROS. The addition of rotenone provides excellent control to this mechanism because this compound blocks the point on which the electrons leak from the Complex I. The last step, the titration of ADP at saturating concentrations to reach OXPHOS allow us to harmonize this protocol with SUIT-026 O2 mt D063 for quality control.
Communicated by Iglesias-Gonzalez J and Gnaiger E (last update 2019-06-26)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
0DTPA |
| |||
0SOD |
| |||
0HRP |
| |||
0AmR |
|
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1S | SL(n) | S | CII | 1S
|
2Rot | SL(n) | S | CII | 1S;2Rot
|
3D | SP | S | CII | 1S;2Rot;3D
|
4Ama | ROX | 1S;2Rot;3D;4Ama
|
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- 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
Compare SUIT protocols
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Fluorometry