通过10BASE(d)-T获得的用于蛋白质特异性指示的变色和强度增强荧光探针的选择。
Selection of Color-Changing and Intensity-Increasing Fluorogenic Probe as Protein-Specific Indicator Obtained via the 10BASE(d)-T.
作者信息
Taki Masumi, Inoue Hiroaki, Mochizuki Kazuto, Yang Jay, Ito Yuji
机构信息
Department of Engineering Science, Bioscience and Technology Program, The Graduate School of Informatics and Engineering, The University of Electro-Communications (UEC) , 1-5-1 Chofugaoka, Chofu, Tokyo 182-8585, Japan.
Department of Anesthesiology, University of Wisconsin, School of Medicine and Public Health , Madison, Wisconsin 53706 United States.
出版信息
Anal Chem. 2016 Jan 19;88(2):1096-9. doi: 10.1021/acs.analchem.5b04687. Epub 2016 Jan 6.
To obtain a molecular probe for specific protein detection, we have synthesized fluorogenic probe library of vast diversity on bacteriophage T7 via the gp10 based-thioetherificaion (10BASE(d)-T). A remarkable color-changing and turning-on probe was selected from the library, and its physicochemical properties upon target-specific binding were obtained. Combination analyses of fluorescence emission titration, isothermal titration calorimetry (ITC), and quantitative saturation-transfer difference (STD) NMR measurements, followed by in silico docking simulation, rationalized the most plausible geometry of the ligand-protein interaction.
为了获得用于特异性蛋白质检测的分子探针，我们通过基于gp10的硫醚化反应（10BASE(d)-T）在噬菌体T7上合成了具有广泛多样性的荧光探针文库。从文库中筛选出了一种显著变色且开启型的探针，并获得了其在特异性结合靶点时的物理化学性质。通过荧光发射滴定、等温滴定量热法（ITC）和定量饱和转移差（STD）核磁共振测量的组合分析，随后进行计算机对接模拟，阐明了配体 - 蛋白质相互作用最合理的几何结构。
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