- high-resolution terminology - matching measurements at high-resolution
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 |
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.
- + 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.
- - 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 (GMP is inhibited by the CII inhibitor malonic acid to a larger extent than PMP). 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 GMP, 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 NSP- and NSE-capacities can only be compared with NP- and SE-capacities. This is not a problem when NSP = NSE (Gnaiger 2009). Otherwise, it may be assumed that SP = SE (Votion et al 2012), such that NSP can be compared with NP + SP. 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-
1GM;2D;3S;3c;4U;5Rot;6Ama
MitoPedia concepts: SUIT protocol, SUIT A
