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Nat Chem Biol. 2017 Sep;13(9):1045-1052. doi: 10.1038/nchembio.2417. Epub 2017 Jun 26.

Monitoring thioredoxin redox with a genetically encoded red fluorescent biosensor.

Author information

1
The Environmental Toxicology Graduate Program, University of California Riverside, Riverside, California, USA.
2
Department of Chemistry, University of California Riverside, Riverside, California, USA.

Abstract

Thioredoxin (Trx) is one of the two major thiol antioxidants, playing essential roles in redox homeostasis and signaling. Despite its importance, there is a lack of methods for monitoring Trx redox dynamics in live cells, hindering a better understanding of physiological and pathological roles of the Trx redox system. In this work, we developed the first genetically encoded fluorescent biosensor for Trx redox by engineering a redox relay between the active-site cysteines of human Trx1 and rxRFP1, a redox-sensitive red fluorescent protein. We used the resultant biosensor-TrxRFP1-to selectively monitor perturbations of Trx redox in various mammalian cell lines. We subcellularly localized TrxRFP1 to image compartmentalized Trx redox changes. We further combined TrxRFP1 with a green fluorescent Grx1-roGFP2 biosensor to simultaneously monitor Trx and glutathione redox dynamics in live cells in response to chemical and physiologically relevant stimuli.

PMID:
28671680
PMCID:
PMC5605834
DOI:
10.1038/nchembio.2417
[Indexed for MEDLINE]
Free PMC Article

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