Difference between revisions of "SUIT-031"
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS ย | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS ย | ||
::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3S;4Rot;5U;- | ::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3S;4Rot;5U;- | ||
This is a protocol to investigate oxygen fluxย [[NADH Electron transfer-pathway state| N-]] and [[NS-pathway control state| NS-pathway]] and [[S-pathway control state]] in the OXPHOS state and [[S-pathway control state]] in the ET state in isolated mitochondria and permeabilized cells. [[SUIT-031 Q mt D072]] and [[SUIT-031 Q ce-pce D074]] are designed to measure simultaneously O<sub>2</sub> flux and the redox state | This is a protocol to investigate oxygen fluxย [[NADH Electron transfer-pathway state| N-]] and [[NS-pathway control state| NS-pathway]] and [[S-pathway control state]] in the OXPHOS state and [[S-pathway control state]] in the ET state in isolated mitochondria and permeabilized cells. [[SUIT-031 Q mt D072]] and [[SUIT-031 Q ce-pce D074]] are designed to measure simultaneously O<sub>2</sub> flux and the Q-redox state in isolated mitochondria and permeabilized cells. ย | ||
''in preparation'' | ''in preparation'' | ||
__TOC__ | __TOC__ |
Revision as of 12:02, 2 December 2021
Description
Abbreviation: PM+S+Rot
Reference: A:
- SUIT-category: NS
- SUIT protocol pattern: orthogonal 1PM;2D;3S;4Rot;5U;-
This is a protocol to investigate oxygen flux N- and NS-pathway and S-pathway control state in the OXPHOS state and S-pathway control state in the ET state in isolated mitochondria and permeabilized cells. SUIT-031 Q mt D072 and SUIT-031 Q ce-pce D074 are designed to measure simultaneously O2 flux and the Q-redox state in isolated mitochondria and permeabilized cells. in preparation
Communicated by Komlodi T (names to be added) (last update 2021-11-25) in preparation
Specific SUIT protocols
SUIT-031 Q mt D072
- SUIT-031 Q mt D072 simultaneous determination of O2 flux and the Q-redox state in isolated mitochondria and cells permeabilized previously to addition to the O2k-chamber.
SUIT-031 Q ce-pce D074
- SUIT-031 Q ce-pce D074 simultaneous determination of O2 flux and the Q-redox state in permeabilized cells.
SUIT-031 O2 mt D075
- SUIT-031 O2 mt D075 for study N-, NS- and S(Rot)- pathways in isolated mitochondria and cells permeabilized previously to addition to the O2k-chamber.
SUIT-031 O2 ce-pce D079
- SUIT-031 O2 ce-pce D079 for study N-, NS- and S(Rot)- pathways in permeabilized cells.
Strengths and limitations
- NS-OXPHOS capacity provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
- A succinate concentration of >10 mM may be required for saturating SE capacity.
- + Reasonable duration of the experiment.
- + You obtain information in a single protocol about the NS-, S and N- pathway control state.
- - CIV activity cannot be determined and Cytochrome c test cannot be performed when measuring the Q redox state.
- - Careful washing is required after the experiment to avoid carry-over of the inhibitors and uncoupler.
Compare SUIT protocols
- [SUIT-001]] is a coupling-control protocol to asses the N-pathway and the contribution of G-, S-, Oct-, and Gp- pathways in the ET state.
- SUIT-006 O2 mt D047 is a coupling-control protocol in the N-pathway for isolated mitochondria, tissue homogenate and cells permeabilized before addition to the O2k- chamber.
- SUIT-004 is designed to provide a quick assessment of the linear coupling control (L- P- E) with NADH-linked substrates (PM) and the contribution of the S-pathway to the ET state (N, NS, S).
- SUIT-008 is designed to assess the additivity between the N- and S-pathway in the Q-junction, providing a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
References
MitoPedia concepts: MiP concept, SUIT protocol
MitoPedia methods:
Respirometry