SUIT-031: Difference between revisions

Latest revision as of 17:57, 13 December 2023

high-resolution terminology - matching measurements at high-resolution

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 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 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-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

@@ Line 1: / Line 1: @@
 {{MitoPedia
-|abbr=PM+S+Rot_OXPHOS
+|abbr=PM+S+Rot
-|description=[[File:1PM;2D;3G;4S;5Rot;6Omy;7U-.png|400px]]
+|description=[[File:1PM;2D;3S;4Rot;5U;-.png|400px]]
-|info='''A: simultaneous determination of O2 flux and the redox state of the Q-pool'''
+|info='''A: '''
 }}
 ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS
 ::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3S;4Rot;5U;-
-This is a protocol to investigate simultanouesly oxygen flux and the redox state of the Q-pool in the [[NADH Electron transfer-pathway state| N-]] and [[NS-pathway control state| NS-pathway]] control state on isolated mitochondria and permeabilized cells.
+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__
-  Communicated by  [[Komlodi T]] (names to be added) (last update 2021-02-01)
+  Communicated by  [[Komlodi T]] (names to be added) (last update 2021-11-25) ''in preparation''
 == Specific SUIT protocols ==
 === SUIT-031 Q mt D072 ===
-[[File:1PM;2D;2S;4Rot;5U;6Ama.png|300px]]
+[[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 Q.png|600px]]
-:::* [[SUIT-031 Q mt D072]]  simultaneous determination of O<sub>2</sub> flux and the redox ratio of the Q-pool in isolated mitochondria.
+:::* [[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;2S;4Rot;5U;6Ama.png|300px]]
+[[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 Q.png|600px]]
-:::* [[SUIT-031 Q ce-pce D074]] simultaneous determination of O<sub>2</sub> flux and the redox ratio of the Q-pool in permeabilized cells.
+:::* [[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;2S;4Rot;5U;6Ama.png|300px]]
+[[File:1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png|300px]]
-[[File:SUIT-031 O2 mt D075.png|600px]]
+[[File:D075 O2 traces.png||600px]]
-{{Template:SUIT-031}}
+:::* [[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 ===
+[[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 ==
 :::* 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.
+:::* 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 together with the fluorescence dyes.
+:::- 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.
@@ Line 55: / Line 62: @@
 }}
-{{#ask:[[Category:Abstracts]]  [[Additional label::SUIT-031]] [[Instrument and method::O2k-Protocol]]
+{{#ask:[[Category:Abstracts]]  [[Additional label::SUIT-031 Q mt D072]] [[Instrument and method::O2k-Protocol]]
 |?Was submitted in year=Year
 |?Has title=Reference
@@ Line 68: / Line 75: @@
 {{MitoPedia concepts
-|mitopedia concept=MiP concept, SUIT protocol, Recommended
+|mitopedia concept=MiP concept, SUIT protocol
 }}
 {{MitoPedia methods
 |mitopedia method=Respirometry,
 }}