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Doerrier 2016 Abstract MitoFit Science Camp 2016

From Bioblast
SUIT reference assay for OXPHOS analysis by high-resolution respirometry.

Link:

Doerrier C, Sumbalova Z, Lamberti G, Krumschnabel G, Gnaiger E (2016)

Event: MitoFit Science Camp 2016 Kuehtai AT

A substrate-uncoupler-inhibitor titration (SUIT) reference assay (SUIT-RA) has been developed to provide a common baseline for comparison of mitochondrial respiratory control in a large variety of species, tissues and cell types, mt-preparations and laboratories, for establishing a database on comparative mitochondrial phyisology. The SUIT-RA is applied in the MitoFit proficiency test with HEK 293T cells. It includes a large number of chemicals used in various specific SUIT protocols, subjecting these chemicals under quality control in the MitoFit proficiency test. The SUIT-RA consists of two coordinated SUIT reference protocols (SUIT-RP). The SUIT-RP1 (RP1) allows to evaluate CI-linked linear coupling control: L – P - E, thus separating coupling control (CI-linked) and substrate control (in the ETS state) (Figure 1). The SUIT-RP2 (RP2) provides information about fatty acid oxidation (FAO) FAOP compared to CI&FAOP. Moreover, RP2 permits to measure maximum ETS capacity through CI, from FAO oxidation through electron-transferring flavoprotein complex (CETF), from succinate (S) through CII, from glycoreophosphate (Gp) through glycerophosphate dehydrogenase (GpDH) on the pathway level of converging into CoQ: PGMSOctGpE (Figure 2). RP1 and RP2, both supply information downstream CoQ, giving an evaluation of CIV activity. These SUIT-RP are harmonized (Figure 3) such that they can be statistically evaluated as replicate measurement for carefully selected respiratory states, while additional information is obtained when the two protocols are conducted in parallel. Therefore, the two SUIT-RP are complementary with their focus on specific respiratory coupling and substrate control aspects, extending previous strategies for respirometrc OXPHOS analysis.


β€’ O2k-Network Lab: AT Innsbruck OROBOROS, AT Innsbruck Gnaiger E


Labels: MiParea: Respiration 


Organism: Human  Tissue;cell: Kidney 


Coupling state: LEAK, OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. 

HRR: Oxygraph-2k  Event: A1  MitoFit Science Camp 2016 

Affiliations

1-OROBOROS INSTRUMENTS, Innsbruck; 2-D. Swarovski Research Lab, Dept Visceral, Transplant and Thoracic Surgery, Innsbruck Med Univ, Austria. - [email protected]

Figure

MitoFit Training Camp 2016 Doerrier Figure1.jpg
MitoFit Training Camp 2016 Doerrier Figure2.jpg
MitoFit Training Camp 2016 Doerrier Figure3.jpg

Support

The project was supported by K-Regio project MitoFit, funded in part by the Tyrolian Government and the European Regional Development Fund (ERDF).