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| {{MitoPedia
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| |abbr=RP2
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| |description=[[File:1D;2M.1;3Oct;4M2;5P;6G;7S;8Gp;9U;10Rot-.png|400px]]
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| |info='''A:''' [[Doerrier 2018 Methods Mol Biol]]
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| }}
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| ::: '''[[SUIT protocol pattern]]:''' 1D;2M;3Oct;4M;5P;6G;7S;8Gp;9U;10Rot-
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| (Page under construction) The SUIT-002 protocol in combination with [[SUIT-001]] provides a common reference for comparison of respiratory control of [[Mitochondrial preparations| mitochondrial preparations]] in a wide variety of species, tissues and cell types. SUIT-002 is specially designed to give information on [[Fatty_acid_oxidation_pathway_control_state|F-pathway]] in [[Oxidative phosphorylation|OXPHOS state]] avoiding FAO overestimation in the presence of [[Anaplerosis|anaplerotic]] pathways. Moreover, the pathway control in OXPHOS state ([[Fatty_acid_oxidation_pathway_control_state|F]], F(N), [[FN]], [[FNS]], [[FNSGp]] pathways) and in [[ET capacity| ET state]] ([[FNSGp]] and [[SGp-pathway control state|SGp]]) can be evaluated by using this SUIT protocol. SUIT-002 can be extended with the CIV assay module.
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| __TOC__
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| Communicated by [[Doerrier C]] and [[Gnaiger E]] (last update 2019-06-05)
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| == Specific SUIT protocols ==
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| === SUIT-002 O2 mt D005 and SUIT-002 O2 pfi D006 ===
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| [[File:1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd.png|400px]]
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| [[File:D005_O2_traces.png|400px]]
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| * [[SUIT-002 O2 mt D005]] for [[Isolated mitochondria|isolated mitochondria]], [[Tissue homogenate|tissue homogenate]] and [[Permeabilized cells|permeabilized cells]] (already permeabilized when they are added to the chamber)
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| * [[SUIT-002 O2 pfi D006]] for [[Permeabilized_muscle_fibers|permeabilized muscle fibers]]
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| === SUIT-002 O2 ce-pce D007 ===
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| [[File:ce1;1Dig;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd.png|400px]]
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| [[File:D07_O2_traces.png|400px]]
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| * [[SUIT-002 O2 ce-pce D007]] for [[Living cells|living cells]] (ce) and [[Permeabilized cells|permeabilized cells]] (pce; permeabilization of the plasma membrane occurs into the chambers through digitonin addition)
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| === SUIT-002 O2 ce-pce D007a ===
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| [[File:ce1;1Dig;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd.png|400px]]
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| [[File:D07a_O2_traces.png|400px]]
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| * [[SUIT-002 O2 ce-pce D007a]] for [[Living cells|living cells]] (ce) and [[Permeabilized cells|permeabilized PBMC and PLT]] (pce; permeabilization of the plasma membrane occurs into the chambers through digitonin addition)
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| {{Template:SUIT-002}}
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| == Strengths and limitations ==
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| :::* SUIT-002 in combination with [[SUIT-001]] provides a common reference for comparison of respiratory control in a large variety of species, tissues and cell types. Both SUIT protocols provide a mitochondrial mapping which allows:
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| :::: 1. to obtain reference values.
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| :::: 2. to evaluate mitochondrial physiological diversity, generating a mt-database on comparative mitochondrial physiology.
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| :::: 3. to screen specific defects.
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| :::* [[SUIT-001]] and SUIT-002 are used in the [[Proficiency_test| MitoFit Proficiency test]] for inter-individual and inter-laboratory reproducibility quality control.
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| :::* A succinate concentration of >10 mM may be required for saturating S<sub>''E''</sub> capacity.
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| :::+ SUIT-002 allows the depletion of endogenous substrates with ADP (1D).
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| :::+ The protocol provides information on F-pathway in OXPHOS state. The low concentration of malate used in this protocol, typically 0.1 mM, does not saturate the N-pathway; but saturates the F-pathway.
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| :::+ F-pathway (3Oct-2M.1) can be compared to FN-pathway (5P) in OXPHOS state.
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| :::+ Pathway control in OXPHOS (F, F(N), FN, FNS, FNSGp pathways) and in ET state (FNSGp and SGp) can be observed.
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| :::+ Harmonization with [[SUIT-001]] allows to perform both SUIT protocols in parallel. The cross-linked respiratory states can be statistically used as repeated measurements.
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| :::+ Harmonization with many SUIT protocols (up to step 7S).
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| :::+ In SUIT-002, the full set of pathways converging into Q (FNSGp) is obtained in OXPHOS and ET states. Therefore, ''P''/''E'' (8Gp/9U) at high ET capacity can be calculated.
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| :::+ This protocol can be extended with the Complex IV module.
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| :::- S-pathway in ET state is not obtained (it is obtained in [[SUIT-001]]).
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| :::- Lengthy duration of the experiment.
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| == Compare SUIT protocols ==
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| ::::* [[SUIT-002 O2 ce-pce D007]] (general SUIT-002 for permeabilized cells) differs from [[SUIT-002 O2 ce-pce D007]] (SUIT-002 for permeabilized PBMC and PLT) in the concentration and volume of the titrations.
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| ::::* [[SUIT-002 O2 ce-pce D007]] for permeabilized cells (ROUTINE respiration in [[Living cells|living cells]] is obtained before selectively permeabilizing the plasma membrane).
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| ::::* [[SUIT-002 O2 ce-pce D007a]] for permeabilized PBMC and PLT (ROUTINE respiration in [[Living cells|living cells]] is obtained before selectively permeabilizing the plasma membrane).
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| ::::* [[SUIT-001]] (RP1): Harmonized SUIT protocol.
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| ::::* [[SUIT-005]]
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| ::::* [[SUIT-025]] SUIT-025 allows the evaluation of different ET-pathway states (F, F(N), FN, FNS and S) in OXPHOS state.
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| ::::* [[SUIT-027]] SUIT-027 focuses on the analysis of the malate-anaplerotic pathway control state in mitochondrial preparations.
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| == References ==
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| {{#ask:[[Category:Publications]] [[Instrument and method::O2k-Protocol]] [[Additional label::SUIT-002]]
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| |?Was published in year=Year
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| |?Has title=Reference
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| |?Mammal and model=Organism
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| |?Tissue and cell=Tissue;cell
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| |format=broadtable
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| |limit=5000
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| |offset=0
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| |sort=Was published in year
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| |order=descending
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| }}
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