Difference between revisions of "SUIT-015"

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SUIT-015

Description

Abbreviation: FNS(Oct,PGM)

Reference: A Schoepf 2016 FEBS J

MitoPedia concepts: SUIT protocol, SUIT B

SUIT-category: FNS(Oct,PGM)

SUIT protocol pattern: diametral 1OctM;2D;3G;4P;5S;6U;7Rot

References

	Year	Reference	Organism	Tissue;cell
Schoepf 2016 FEBS J	2016	Schöpf B, Schäfer G, Weber A, Talasz H, Eder IE, Klocker H, Gnaiger E (2016) Oxidative phosphorylation and mitochondrial function differ between human prostate tissue and cultured cells. https://doi.org/10.1111/febs.13733	Human	Endothelial;epithelial;mesothelial cell Genital Other cell lines Fibroblast

Template:SUIT-0XX

Strengths and limitations

+ The protocol provides information on FAO capacity in the absence of other, potentially interfering pathways, both in the LEAK state and in OXPHOS.

+ FNS OXPHOS capacity comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.

+ FNS ET capacity is a good estimate of overal ET capacity in may cell types.

+ The presence of PMG and S establishes fully operative TCA cycle activity.

+ Application of the cytochrome c test early in the protocol ensures comparability of all states in case of any effect of c.

+ Reasonable duration of the experiment.

- F OXPHOS capacity may be underestimated. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) + malate increased OXPHOS by 26% (Lemuieux et al 2011).

- SRot(E) may be underestimated if S is not saturating.

- CIV activity is not measured, to save experimental time.

Compare SUIT protocols

1GM;2D;3S;3c;4U;5Rot;6Ama

MitoPedia concepts: SUIT protocol, SUIT A

@@ Line 32: / Line 32: @@
 :::+ FNS OXPHOS capacity comprises the most important pathways in many cell types and thus provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
 :::+ FNS ET capacity is a good estimate of overal ET capacity in may cell types.
+:::+ The presence of PMG and S establishes fully operative TCA cycle activity.
 :::+ Application of the cytochrome ''c'' test early in the protocol ensures comparability of all states in case of any effect of ''c''.
 :::+ Reasonable duration of the experiment.
+:::- F OXPHOS capacity may be underestimated. In human heart muscle addition of Oct to palmitoylcarnitine (Pal) + malate increased OXPHOS by 26% (Lemuieux et al 2011).
 :::- SRot(E) may be underestimated if S is not saturating.
 :::- CIV activity is not measured, to save experimental time.
-:::- GM and PM yield typically identical fluxes in human skeletal muscle fibres. However, PM is the superior alternative to GM: the fraction of the N-pathway is lower and of the S-pathway is higher with GM compared to PM (GM<sub>''P''</sub> is inhibited by the CII inhibitor malonic acid to a larger extent than PM<sub>''P''</sub>). PM, therefore, yields a more sensitive assay for the diagnosis of injuries in the N-pathway, since an impairment of N-pathway capacity can be compensated partially by activation of the S-pathway. This is a disadvantage compared to SUIT-004 and SUIT-008 for diagnosis of N-capacity.
-:::- To detect an additive effect of P after GM<sub>''P''</sub>, pyruvate would have to be added as step 3 (before S). However, inhibition of respiration was observed after titration of P (5 mM) in horse skeletal muscle fibres (Votion et al 2012), which was not the case when P was titrated in steps of 1 mM.
-:::- When evaluating the additive effect of the N- and S-pathway, it has to be considered that NS<sub>''P''</sub>- and NS<sub>''E''</sub>-capacities can only be compared with N<sub>''P''</sub>- and S<sub>''E''</sub>-capacities. This is not a problem when NS<sub>''P''</sub> = NS<sub>''E''</sub> (Gnaiger 2009). Otherwise, it may be assumed that S<sub>''P''</sub> = S<sub>''E''</sub> (Votion et al 2012), such that NS<sub>''P''</sub> can be compared with N<sub>''P''</sub> + S<sub>''P''</sub>. SUIT-004 should be chosen for the additive effect in the ET-state.
-:::- ''Rox'' may be lower in substrate states earlier in the SUIT protocol. Therefore, this ''Rox'' measurement is frequently taken as a methodological control rather than as the final basis of ''Rox'' correction of mitochondrial respiration (mt).
-:::- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
 == Compare SUIT protocols ==
-::::* GM and PM yield typically identical fluxes in human skeletal muscle fibres.
+::::*
-::::* [[SUIT-004]] 1PM;2D;3U;4S;5Rot-
-::::* [[SUIT-008]] 1PM;2D;3G;4S;5U;6Rot-
-::::* [[1PM;2D;3U;4G;5S;6Oct;7Rot;8Gp-]]
-::::* [[1PGM;2D;3S;4U;5Rot-]]