Abbreviation: CCP mt PM
SUIT number: D047_1PM;2D;2c;3Omy;4U;5Ama
- SUIT protocol pattern: 1PM;2D;2c;3Omy;4U;5Ama
SUIT-006 O2 mt D047 is designed to study different coupling control states (L(n) - P - L(Omy) - E) at N(PM) pathway control state. The use of oligomycin is optional, however, it provides important information when endogenous adenylates are present and ATPases are active in the sample. This situation may lead to overestimated LEAK respiration measured in the absence of adenylates - L(n). Therefore, oligomycin can be used to verify whether this occurs and obtain the LEAK state appropriately.
Communicated by Cardoso LHD, Iglesias-Gonzalez J, Komlodi T and Gnaiger E (last update 2019-06-28)
Steps and respiratory states
|Step||State||Pathway||Q-junction||Comment - Events (E) and Marks (M)
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- Pathway control
Strengths and limitations
- This protocol is indicated when the aim is to analyse coupling control in mitochondrial preparations in NADH-pathway using PM as substrates.
- + Reasonable duration of the experiment.
- + This protocol can be extended with the Complex IV module.
- - This protocol analyses the coupling control in NADH-pathway (with PM), however, it is less specific than a similar protocol containing the complex II inhibitor malonic acid, that would allow to analyse the NADH-pathway without any contribution of the S-pathway. A DLPU can be created, with the addition of an event for malonic acid right before the addition of the N-linked substrates.
- - Oligomycin may lead to underestimation of ET capacity. This may be prevented by the addition of low concentrations of oligomycin, which should be previously tested.
- - Careful washing is required after the experiment to avoid carry-over of uncoupler and inhibitors, if used.
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MitoPedia methods: Respirometry