首页|Development of tricyanofuran-based activity probes for sulfatase assay in live cells
Development of tricyanofuran-based activity probes for sulfatase assay in live cells
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NSTL
Sulfatase plays a pivotal role in the regulation of biologically active metabolites associated with various diseases.In particular,the up-regulation of steroid sulfatase activity has a positive correlation with breast cancer due to the increment of estrone levels via the desulfation reaction of estron 3-sulfate.Despite the high association between sulfatase activity and disease occurrence,there has been little progress in the discovery of sulfatase inhibitors,possibly due to the lack of robust activity-based assays.To design a sulfatase activity probe,we examined the binding between four fluorophores(coumarin,BODIPY,rhodol,and tricyanofuran[TCF])and three isoforms of sulfatase using docking simulations and found that the rhodol and TCF scaffolds exhibited stable binding.To avoid the charge-driven organelle accumulation bias,we chose TCF as the core structure and designed two probes containing sulfate(TCF-OSulf and TCF-NCOO-OSulf).The TCF probes showed clear colorimetric response and 2.7-fold and 5-fold fluorescence enhancements in vitro.In addition,the probes detected the inhibition of the enzyme by estrone-O-sulfamate(EMATE)in a time-dependent manner.Since cell-based screening has many advantages over the traditional high-throughput assay in vitro,we further examined TCF probes for the inhibitor assay for sulfatase activity in live cells.Based on our observations,TCF-NCOO-OSulf could be used to monitor sulfatase activity in live cells at a concentration of 5 ^iM.This is the first sulfatase activity-based probe that can visualize activity and inhibition in live cell conditions.
NSTL
