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A comparison of Prx- and OxyR-based H 2 O 2 probes expressed in S. cerevisiae.

Authors :: Kritsiligkou P
Shen TK
Dick TP
Source :: The Journal of biological chemistry [J Biol Chem] 2021 Jul; Vol. 297 (1), pp. 100866. Date of Electronic Publication: 2021 Jun 09.
Publication Year :: 2021
Abstract: Genetically encoded fluorescent H <subscript>2</subscript> O <subscript>2</subscript> probes continue to advance the field of redox biology. Here, we compare the previously established peroxiredoxin-based H <subscript>2</subscript> O <subscript>2</subscript> probe roGFP2-Tsa2ΔC <subscript>R</subscript> with the newly described OxyR-based H <subscript>2</subscript> O <subscript>2</subscript> probe HyPer7, using yeast as the model system. Although not as sensitive as roGFP2-Tsa2ΔC <subscript>R</subscript> , HyPer7 is much improved relative to earlier HyPer versions, most notably by ratiometric pH stability. The most striking difference between the two probes is the dynamics of intracellular probe reduction. HyPer7 is rapidly reduced, predominantly by the thioredoxin system, whereas roGFP2-Tsa2ΔC <subscript>R</subscript> is reduced more slowly, predominantly by the glutathione system. We discuss the pros and cons of each probe and suggest that future side-by-side measurements with both probes may provide information on the relative activity of the two major cellular reducing systems.<br />Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.<br /> (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)