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Selection of Color-Changing and Intensity-Increasing Fluorogenic Probe as Protein-Specific Indicator Obtained via the 10BASE d -T

2016; American Chemical Society; Volume: 88; Issue: 2 Linguagem: Inglês

10.1021/acs.analchem.5b04687

1520-6882

Masumi Taki, Hiroaki Inoue, Kazuto Mochizuki, Jay Yang, Yuji Ito,

Bacteriophages and microbial interactions

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.