Difference between revisions of "SUIT-022"

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{{MitoPedia
 
{{MitoPedia
|info='''A: Determination of the respiration due to the alternative oxidase pathway''' [[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/6/64/SUIT-022.pdf|Bioblast pdf]] »[http://wiki.oroboros.at/index.php/File:SUIT-022.pdf Versions]
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|abbr=AOX (ce CN+SHAM)
|application=O2
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|description= [[File:ce1;ce2KCN;ce3SHAM.v2.png|400px]]
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|info='''A: Determination of the respiration due to the alternative oxidase pathway'''
 
}}
 
}}
::: '''[[Categories of SUIT protocols |SUIT-category]]:''' ce
+
::: '''[[SUIT protocol pattern]]:''' ce1;ce2KCN;ce3SHAM
::: '''[[SUIT protocol pattern]]:''' linear ce1;ce2KCN;ce3SHAM
 
 
 
The SUIT-022 is designed to provide an estimation of the mitochondrial respiration in cells due to the [[alternative oxidase]] pathway, which is [[cyanide| cyanide]]-and [[Antimycin_A| antimycin A]]-resistant. This protocol must be complemented with the study in parallel of the mitochondrial respiration due to the cytochrome ''c'' pathway (see [[SUIT-023]]).
 
  
 +
The SUIT-022 provides an estimation of mitochondrial respiration in cells through the Q-[[alternative oxidase]] pathway (Q-AOX), which is [[Antimycin_A| antimycin A]]- and [[cyanide| cyanide]] resistant. This protocol must be complemented with the study in parallel of mitochondrial respiration due to the CIII-[[Complex_IV| cytochrome ''c'' oxidase]] pathway (CIII-CIV; see [[SUIT-023]]).
  
 
__TOC__
 
__TOC__
  Communicated by [[Huete-Ortega M]] and [[Gnaiger E]] (last update 2019-04-09)
+
  Communicated by [[Huete-Ortega M]], [[Gnaiger E]] (last update 2019-06-05)
  
 
== Specific SUIT protocols ==
 
== Specific SUIT protocols ==
  
 
=== SUIT-022 O2 ce D051 ===
 
=== SUIT-022 O2 ce D051 ===
[[File:ce1;ce2KCN;ce3SHAM.png|400px]]
+
[[File:ce1;ce2KCN;ce3SHAM.v2.png|400px]] [[File:ce1;ce2KCN;ce3SHAM.png|400px]]
 
[[File:D051_O2_traces.png|400px]]
 
[[File:D051_O2_traces.png|400px]]
*  [[SUIT-022 O2 ce D051]] for intact microalgal cells (ce)
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*  [[SUIT-022 O2 ce D051]] for living microalgal cells (ce)
  
{{Template:SUIT-022}}
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{{Template:SUIT-022 O2 ce D051}}
  
 
== Strengths and limitations ==
 
== Strengths and limitations ==
  
:::* SUIT-022 provides an estimation of the [[cyanide| cyanide]]-and [[Antimycin_A| antimycin A]]-resistant mitochondrial respiration due to the activity of the [[alternative oxidase]] on cells on [[ROUTINE]]. To complement the study with the estimation of the mitochondrial respiration due to the cytochrome ''c'' pathway, it must be carried out in parallel to [[SUIT-023]].
+
:::* SUIT-022 provides an estimation of the [[cyanide| cyanide]]-and [[Antimycin_A| antimycin A]]-resistant mitochondrial respiration due to the activity of [[alternative oxidase]] in cells on [[ROUTINE]]. To complement the study with the estimation of mitochondrial respiration due to the CIII-CIV pathway, it must be carried out in parallel to [[SUIT-023]].
:::+ This protocol can be used with many type of cells, although optimal concentrations of inhibitors have to be optimised.
+
:::+ This protocol can be used with many types of cells, although optimal concentrations of inhibitors have to be determined.
 
:::+ Short duration of experiment
 
:::+ Short duration of experiment
  
:::- Estimation of the linear coupling control is not done
+
:::- No estimation of coupling control
:::- The use of [[cyanide| cyanide]] requires a careful and in deep cleaning of the chambers after use and careful handling of the chemical to guarantee health safety.
+
:::- The use of [[cyanide| cyanide]] requires careful cleaning of the chambers after use and careful handling of the chemical to guarantee health safety.
 +
 
  
 
== Compare SUIT protocols ==
 
== Compare SUIT protocols ==
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|mitopedia method=Respirometry
 
|mitopedia method=Respirometry
 
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{{MitoPedia O2k and high-resolution respirometry}}
 
{{MitoPedia topics}}
 

Latest revision as of 08:53, 15 June 2019

Bioblasts - Richard Altmann and MiPArt by Odra Noel
MitoPedia         Terms and abbreviations         Concepts and methods         MitoPedia: SUIT         MiP and biochemistry         Preprints and history



MitoPedia

SUIT-022

Description

Ce1;ce2KCN;ce3SHAM.v2.png

Abbreviation: AOX (ce CN+SHAM)

Reference: A: Determination of the respiration due to the alternative oxidase pathway

SUIT protocol pattern: ce1;ce2KCN;ce3SHAM

The SUIT-022 provides an estimation of mitochondrial respiration in cells through the Q-alternative oxidase pathway (Q-AOX), which is antimycin A- and cyanide resistant. This protocol must be complemented with the study in parallel of mitochondrial respiration due to the CIII- cytochrome c oxidase pathway (CIII-CIV; see SUIT-023).

Communicated by Huete-Ortega M, Gnaiger E (last update 2019-06-05)

Specific SUIT protocols

SUIT-022 O2 ce D051

Ce1;ce2KCN;ce3SHAM.v2.png Ce1;ce2KCN;ce3SHAM.png D051 O2 traces.png

MitoPedia: SUIT

Steps and respiratory states

Ce1;ce2KCN;ce3SHAM.v2.png Ce1;ce2KCN;ce3SHAM.png

Step State Pathway Q-junction Comment - Events (E) and Marks (M)
ce1 ROUTINE ce1
  • ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
ce2KCN ROUTINE or ROX ce1;ce2KCN
  • Cyanide (usually added as KCN) is a competitive inhibitor of cytochrome c oxidase (CIV). In the presence of an active alternative oxidase (AOX) pathway, after the inhibition of CIV, cells remain in the physiological coupling state ROUTINE. In those occasions where the AOX is not active, after the inhibition of CIV, the (ROX state) is normally achieved, where Rox is the residual oxygen consumption due to oxidative side reactions estimated either after inhibition of CIII, CIV or in the absence of endogenous fuel-substrates. Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration. Cyanide inhibition is reversed by pyruvate and high oxygen levels.
ce3SHAM ROX ce1;ce2KCN;ce3SHAM


Questions.jpg


Click to expand or collaps

Strengths and limitations

  • SUIT-022 provides an estimation of the cyanide-and antimycin A-resistant mitochondrial respiration due to the activity of alternative oxidase in cells on ROUTINE. To complement the study with the estimation of mitochondrial respiration due to the CIII-CIV pathway, it must be carried out in parallel to SUIT-023.
+ This protocol can be used with many types of cells, although optimal concentrations of inhibitors have to be determined.
+ Short duration of experiment
- No estimation of coupling control
- The use of cyanide requires careful cleaning of the chambers after use and careful handling of the chemical to guarantee health safety.


Compare SUIT protocols

References

MitoPedia concepts: MiP concept, SUIT protocol, Recommended 


MitoPedia methods: Respirometry