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SUIT-031

From Bioblast


high-resolution terminology - matching measurements at high-resolution


SUIT-031

Description

1PM;2D;3S;4Rot;5U;-.png

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

Mt;1Q2;1PM;2D;3S;4Rot;5U;6Anox;7Ama.png SUIT-031 Q mt D072 O2.png SUIT-031 Q mt D072 Q.png

  • 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

Ce1;1Dig;1Q2;1PM;2D;3S;4Rot;5U;6Anox;7Ama.png SUIT-031 Q ce-pce D074 O2.png SUIT-031 Q ce-pce D074 Q.png

SUIT-031 O2 mt D075

1PM;2D;3S;4Rot;5U;5Anoxia;6Ama.png D075 O2 traces.png

  • 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

Ce1;1Dig;1PM;2D;2c;3S;4Rot;5U;6Ama.png D079 O2 traces.png

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