Making the invisible visible – Mechanistical insights on biosensor functioning

Description

Hypocrates and HyPer7 are fluorescent protein-based biosensors for hypohalous acids and hydrogen peroxide, which are being used by the redox community in a wide variety of experimental settings. Nevertheless, many mechanistic details about the functioning of these biosensors remain unexplored. It is not clear which of the cellular reducing systems - the glutathione/glutaredoxin, or the thioredoxin system, reduces the biosensors. Further, it is not understood how the conformational coupling between the sensing moiety, and the chromophore of cpYFP occurs. Using a combination of X-ray crystallography, artificial intelligence structure predictions, and various biophysical and biochemical techniques I will address these questions and mechanistically dissect Hypocrates and HyPer7. These results will on the one hand contribute to a more informed interpretation and planning of experiments with these biosensors, and on the other hand serve as a basis for the development of other tools to probe for subcellular oxidative and metabolic events.

1 Kostyuk, A.I. et al. (2022) Nature Communications 13,171;
2 Pak, V.V. et al. (2020) Cell Metabolism 31,642;

Period	9 Oct 2022
Event title	EMBO workshop: Thiol oxidation in biology
Event type	Workshop
Location	San Felui, Spain
Degree of Recognition	International