Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, College of Chemistry, Central China Normal University , Wuhan 430079, P.R. China.
Collaborative Innovation Center of Chemical Science and Engineering, Tianjin 30071, P.R. China.
Anal Chem. 2017 Mar 21;89(6):3687-3693. doi: 10.1021/acs.analchem.6b05115. Epub 2017 Mar 7.
We report herein a nonpeptide-based small-molecule probe for fluorogenic and chromogenic detection of chymotrypsin, as well as the primary application for this probe. This probe was rationally designed by mimicking the peptide substrate and optimized by adjusting the recognition group. The refined probe 2 exhibits good specificity toward chymotrypsin, producing about 25-fold higher enhancement in both the fluorescence intensity and absorbance upon the catalysis by chymotrypsin. Compared with the most widely used peptide substrate (AMC-FPAA-Suc) of chymotrypsin, probe 2 shows about 5-fold higher binding affinity and comparable catalytical efficiency against chymotrypsin. Furthermore, it was successfully applied for the inhibitor characterization. To the best of our knowledge, probe 2 is the first nonpeptide-based small-molecule probe for chymotrypsin, with the advantages of simple structure and high sensitivity compared to the widely used peptide-based substrates. This small-molecule probe is expected to be a useful molecular tool for drug discovery and chymotrypsin-related disease diagnosis.
我们在此报告了一种基于非肽的小分子探针，用于糜蛋白酶的荧光和比色检测，以及该探针的初步应用。该探针通过模拟肽底物进行合理设计，并通过调整识别基团进行优化。经过优化的探针 2 对糜蛋白酶具有良好的特异性，在糜蛋白酶催化作用下，荧光强度和吸光度分别提高了约 25 倍。与最广泛使用的糜蛋白酶肽底物 (AMC-FPAA-Suc) 相比，探针 2 对糜蛋白酶的结合亲和力约高 5 倍，催化效率相当。此外，它还成功地应用于抑制剂的表征。据我们所知，探针 2 是第一个基于非肽的小分子糜蛋白酶探针，与广泛使用的基于肽的底物相比，具有结构简单、灵敏度高的优点。这种小分子探针有望成为药物发现和与糜蛋白酶相关疾病诊断的有用分子工具。
