Validation of a respiratory phenotyping assay for yeast indicates that aerobic metabolism is regulated by mitochondrial thioredoxin activity (572.1)

Thiol oxidoreductase enzymes, including thioredoxins, glutaredoxins, peroxiredoxins, and glutathione peroxiredoxins, play an important role in regulating cellular redox reactions through reduction of oxidized thiols. These enzymes regulate integral processes such as protein folding, detoxification of reactive oxygen species, and metal and sulfur metabolism; however, many thiol oxidoreductase substrates and consequential functional changes are not known. Since thiol oxidoreductase genes are highly conserved across eukaryotic species, the objective of this study was to screen a Saccharomyces cerevisiae thiol oxidoreductase deletion library for respiratory phenotypes. A novel protocol using the XF Flux Analyzer (Seahorse Biosciences) was established to simultaneously measure real-time oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) from media which respectively correlates to cellular aerobic and anaerobic respiratory activities in S. cerevisiae. This approach was validated by artifi...