A fast and specific fluorescent probe for thioredoxin reductase that works via disulphide bond cleavage

  • Nat Commun. 2019 Jun 21;10(1):2745. doi: 10.1038/s41467-019-10807-8.
Xinming Li  1 ,  Baoxin Zhang  1 ,  Chaoxian Yan  1 ,  Jin Li  1 ,  Song Wang  1 ,  Xiangxu Wei  1 ,  Xiaoyan Jiang  1 ,  Panpan Zhou  1 ,  Jianguo Fang  2
Affiliations
  • 1. State Key Laboratory of Applied Organic Chemistry & College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China.
  • 2. State Key Laboratory of Applied Organic Chemistry & College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China. [email protected].
PMID: 31227705 DOI: 10.1038/s41467-019-10807-8
Abstract

Small molecule probes are indispensable tools to explore diverse cellular events. However, finding a specific probe of a target remains a high challenge. Here we report the discovery of Fast-TRFS, a specific and superfast fluorogenic probe of mammalian thioredoxin reductase, a ubiquitous enzyme involved in regulation of diverse cellular redox signaling pathways. By systematically examining the processes of fluorophore release and reduction of cyclic disulfides/diselenides by the enzyme, structural factors that determine the response rate and specificity of the probe are disclosed. Mechanistic studies reveal that the fluorescence signal is switched on by a simple reduction of the disulfide bond within the probe, which is in stark contrast to the sensing mechanism of published probes. The favorable properties of Fast-TRFS enable development of a high-throughput screening assay to discover inhibitors of thioredoxin reductase by using crude tissue extracts as a source of the enzyme.

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