Difference between revisions of "SUIT-031"

Revision as of 16:57, 25 November 2021

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SUIT-031

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 redox state of the Q-pool in isolated mitochondria and permeabilized cells.

Communicated by  Komlodi T (names to be added) (last update 2021-11-25)

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

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 ce-pce D000

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 S_E 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-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

@@ Line 1: / Line 1: @@
+{{MitoPedia
+|abbr=PM+S+Rot
+|description=[[File:1PM;2D;3S;4Rot;5U;-.png|400px]]
+|info='''A: '''
+}}
+::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS
+::: '''[[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 of the Q-pool in isolated mitochondria and permeabilized cells.
+__TOC__
+ Communicated by  [[Komlodi T]] (names to be added) (last update 2021-11-25)
+== Specific SUIT protocols ==
+=== SUIT-031 Q mt D072 ===
+[[File:1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png|300px]]
+[[File:SUIT-031 Q mt D072 O2.png|600px]]
+[[File:SUIT-031 Q mt D072 Q.png|600px]]
+:::* [[SUIT-031 Q mt D072]]  simultaneous determination of O<sub>2</sub> 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 ===
+[[File:ce1;1Dig;1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png|400px]]
+[[File:SUIT-031 Q ce-pce D074 O2.png|600px]]
+[[File:SUIT-031 Q ce-pce D074 Q.png|600px]]
+:::* [[SUIT-031 Q ce-pce D074]] simultaneous determination of O<sub>2</sub> flux and the Q-redox state in permeabilized cells.
+=== SUIT-031 O2 mt D075 ===
+[[File:1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png|300px]]
+[[File:SUIT-031 O2 mt D075 O2.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 ce-pce D000 ===
+== 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 S<sub>E</sub> capacity.
+:::+ Reasonable duration of the experiment.
+:::+ You obtain information in a single protocol about the [[NS-pathway control state| NS-]], [[Succinate pathway control state| S]] and [[NADH Electron transfer-pathway state| 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-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 ==
+{{#ask:[[Category:Publications]] [[Instrument and method::O2k-Protocol]] [[Additional label::SUIT-031]]
+|?Was published in year=Year
+|?Has title=Reference
+|?Mammal and model=Organism
+|?Tissue and cell=Tissue;cell
+|format=broadtable
+|limit=5000
+|offset=0
+|sort=Was published in year
+|order=descending
+}}
+{{#ask:[[Category:Abstracts]]  [[Additional label::SUIT-031 Q mt D072]] [[Instrument and method::O2k-Protocol]]
+|?Was submitted in year=Year
+|?Has title=Reference
+|?Mammal and model=Organism
+|?Tissue and cell=Tissue;cell
+|format=broadtable
+|limit=5000
+|offset=0
+|sort=Was published in year
+|order=descending
+}}
+{{MitoPedia concepts
+|mitopedia concept=MiP concept, SUIT protocol
+}}
+{{MitoPedia methods
+|mitopedia method=Respirometry,
+}}