1362851-21-5Relevant articles and documents
A new FRET probe for ratiometric fluorescence detecting mitochondria-localized drug activation and imaging endogenous hydroxyl radicals in zebrafish
Chen, Tongkai,Deng, Tao,Hu, Shiyou,Liu, Fang,Liu, Wei,Wang, Xiaojuan,Wu, Shengjun,Xu, Qin,Yu, Zhiqiang
supporting information, p. 4432 - 4435 (2020/05/05)
A new FRET probe has been prepared for ratiometric fluorescence detection of hydroxyl radicals. It has been successfully used for detecting mitochondria-localized drug activation in living cells and imaging endogenous hydroxyl radicals in zebrafish gastrointestinal (GI) tracts under normal culturing conditions.
Fluorescent Probes for Single-Step Detection and Proteomic Profiling of Histone Deacetylases
Xie, Yusheng,Ge, Jingyan,Lei, Haipeng,Peng, Bo,Zhang, Huatang,Wang, Danyang,Pan, Sijun,Chen, Ganchao,Chen, Lanfang,Wang, Yi,Hao, Quan,Yao, Shao Q.,Sun, Hongyan
supporting information, p. 15596 - 15604 (2016/12/16)
Histone deacetylases (HDACs) play important roles in regulating various physiological and pathological processes. Developing fluorescent probes capable of detecting HDAC activity can help further elucidate the roles of HDACs in biology. In this study, we first developed a set of activity-based fluorescent probes by incorporating the Kac residue and the O-NBD group. Upon enzymatic removal of the acetyl group in the Kac residue, the released free amine reacted intramolecularly with the O-NBD moiety, resulting in turn-on fluorescence. These designed probes are capable of detecting HDAC activity in a continuous fashion, thereby eliminating the extra step of fluorescence development. Remarkably, the amount of turn-on fluorescence can be as high as 50-fold, which is superior to the existing one-step HDAC fluorescent probes. Inhibition experiments further proved that the probes can serve as useful tools for screening HDAC inhibitors. Building on these results, we moved on and designed a dual-purpose fluorescent probe by introducing a diazirine photo-cross-linker into the probe. The resulting probe was not only capable of reporting enzymatic activity but also able to directly identify and capture the protein targets from the complex cellular environment. By combining a fluorometric method and in-gel fluorescence scanning technique, we found that epigenetic readers and erasers can be readily identified and differentiated using a single probe. This is not achievable with traditional photoaffinity probes. In light of the prominent properties and the diverse functions of this newly developed probe, we envision that it can provide a robust tool for functional analysis of HDACs and facilitate future drug discovery in epigenetics.
Activity-based proteome profiling of potential cellular targets of orlistat - An FDA-approved drug with anti-tumor activities
Yang, Peng-Yu,Liu, Kai,Ngai, Mun Hong,Lear, Martin J.,Wenk, Markus R.,Yao, Shao Q.
supporting information; scheme or table, p. 656 - 666 (2010/03/25)
Orlistat, or tetrahydrolipstatin (THL), is an FDA-approved antiobesity drug with potential antitumor activities. Cellular off-targets and potential side effects of Orlistat in cancer therapies, however, have not been extensively explored thus far. In this study, we report the total of synthesis of THL-like protein-reactive probes, in which extremely conservative modifications (i.e., an alkyne handle) were introduced in the parental THL structure to maintain the native biological properties of Orlistat, while providing the necessary functionality for target identification via the bio-orthogonal click chemistry. With these natural productlike, cell-permeable probes, we were able to demonstrate, for the first time, this chemical proteomic approach is suitable for the identification of previously unknown cellular targets of Orlistat. In addition to the expected fatty acid synthase (FAS), we identified a total of eight new targets, some of which were further validated by experiments including Western blotting, recombinant protein expression, and site-directed mutagenesis. Our findings have important implications in the consideration of Orlistat as a potential anticancer drug at its early stages of development for cancer therapy. Our strategy should be broadly useful for off-target identification against quite a number of existing drugs and/or candidates, which are also covalent modifiers of their biological targets.
