Fluorogenic and chromogenic detection of carboxypeptidase Y with a nonpeptide-based small-molecule probe
作者： Hao Xiong , Jia-Qian Yang , Wei-Ming Kang , Feng-Xu Wu , Jia-Li Zuo
DOI: 10.1016/J.SNB.2018.04.102
关键词: Substrate (chemistry) 、 Biochemistry 、 Peptide 、 Chromogenic 、 Small molecule 、 Fluorescence 、 Chemistry 、 Enzyme 、 Amino acid 、 Protein sequencing
摘要: Abstract Carboxypeptidase Y (CPY) is a vital enzyme with many well-known biological significances, such as determination of amino acid sequences and biosynthesis polypeptides. Herein we developed nonpeptide-based fluorescent probe to sense CPY activity accurately successfully applied it the inhibitory study. Based on organic synthesis comprehensive characterization, refined CPY-P3 displayed excellent specificity towards exhibited over 400-fold enhancement fluorescence intensity upon hydrolysis CPY. Compared commonly used peptide-based substrate (Suc-IIW-AMC), showed about 12-fold higher binding affinity comparable catalytical efficiency for Moreover, was in inhibitor characterization. To our knowledge, first small-molecule time-course detection. Meanwhile, generation kelly color makes detection directly visible without any complicated device. In all, this should be potentially useful tool understanding roles protein sequencing engineering, well drug discovery CPY-related diseases.
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