SUIT-031: Difference between revisions
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS | ||
::: '''[[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 flux [[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 flux [[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. | ||
''in preparation'' | |||
__TOC__ | __TOC__ | ||
Communicated by [[Komlodi T]] (names to be added) (last update 2021-11-25) | Communicated by [[Komlodi T]] (names to be added) (last update 2021-11-25) ''in preparation'' | ||
== Specific SUIT protocols == | == Specific SUIT protocols == | ||
=== SUIT-031 Q mt D072 === | === SUIT-031 Q mt D072 === | ||
[[File:1PM;2D;3S;4Rot;5U; | [[File:mt;1Q2;1PM;2D;3S;4Rot;5U;6Anox;7Ama.png|300px]] | ||
[[File:SUIT-031 Q mt D072 O2.png|600px]] | [[File:SUIT-031 Q mt D072 O2.png|600px]] | ||
[[File:SUIT-031 Q mt D072 Q.png|600px]] | [[File:SUIT-031 Q mt D072 Q.png|600px]] | ||
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=== SUIT-031 Q ce-pce D074 === | === SUIT-031 Q ce-pce D074 === | ||
[[File:ce1;1Dig;1PM;2D;3S;4Rot;5U; | [[File:ce1;1Dig;1Q2;1PM;2D;3S;4Rot;5U;6Anox;7Ama.png|400px]] | ||
[[File:SUIT-031 Q ce-pce D074 O2.png|600px]] | [[File:SUIT-031 Q ce-pce D074 O2.png|600px]] | ||
[[File:SUIT-031 Q ce-pce D074 Q.png|600px]] | [[File:SUIT-031 Q ce-pce D074 Q.png|600px]] | ||
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=== SUIT-031 O2 mt D075 === | === SUIT-031 O2 mt D075 === | ||
[[File:1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png|300px]] | [[File:1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png|300px]] | ||
[[File: | [[File:D075 O2 traces.png||600px]] | ||
:::* [[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 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 | === SUIT-031 O2 ce-pce D079 === | ||
[[File:ce1;1Dig;1PM;2D;2c;3S;4Rot;5U;6Ama.png|300px]] | |||
[[File:D079 O2 traces.png||600px]] | |||
:::* [[SUIT-031 O2 ce-pce D079]] for study N-, NS- and S(Rot)- pathways in permeabilized cells. | |||
== Strengths and limitations == | == Strengths and limitations == | ||
:::* NS-OXPHOS capacity provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity. | :::* NS-OXPHOS capacity provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity. | ||
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== Compare SUIT protocols == | == 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-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-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). |
Latest revision as of 17:57, 13 December 2023
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