Difference between revisions of "SUIT-026 AmR mt D064"
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== Strengths and limitations == | == Strengths and limitations == | ||
:::* The conditions under RET is observable in isolated mitochondria are not physiological but it has been suggested that corresponds to some pathological states. | |||
:::+ This protocol must be run parallel with SUIT-026 O2 mt D063 to have a respiratory control. | |||
:::+ Short protocol. | |||
:::+ Rotenone provides an excellent control step for RET. | |||
:::- The protocol does not allow to obtain all the coupling states of the sample. | |||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
::::* [[SUIT-006 O2 mt D022]]: CCP mtprep for S-pathway. | |||
::::* [[SUIT-026 O2 mt D063]]: Respiratory control protocol.. | |||
Revision as of 12:48, 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-006 O2 mt D022 for quality control and a full assessment of the coupling 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
|
- 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 conditions under RET is observable in isolated mitochondria are not physiological but it has been suggested that corresponds to some pathological states.
- + This protocol must be run parallel with SUIT-026 O2 mt D063 to have a respiratory control.
- + Short protocol.
- + Rotenone provides an excellent control step for RET.
- - The protocol does not allow to obtain all the coupling states of the sample.
Compare SUIT protocols
- SUIT-006 O2 mt D022: CCP mtprep for S-pathway.
- SUIT-026 O2 mt D063: Respiratory control protocol..
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Fluorometry