期刊
ANALYTICAL CHEMISTRY
卷 85, 期 16, 页码 7957-7965出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac401732d
关键词
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资金
- National Natural Science Foundation of China [31170782]
- Tianjin Natural Science Foundation [11JCYBJC25500]
- Ph.D. Programs Foundation of the Ministry of Education of China [20110031120019]
- National Biomedical Special Project of International Innovation Park [11ZCKFSY06300]
Mass spectrometry-based platforms have gained increasing success in discovery of ligands bound to therapeutic targets as drug candidates. We established both a nanoelectrospray ionization mass spectrometry (nanoESI-MS) assay and an ultrafiltration liquid chromatography/mass spectrometry (LC/MS) assay to identify new ligands for New Delhi metallo-beta-lactamase 1 (NDM-1), responsible for worldwide antibiotic resistance. To alleviate nonspecific binding of hydrophobic compounds and eliminate false positives typically encountered in the indirect LC/MS-based assay, we introduced a blocking protein in the control, which remarkably enhances the selectivity and accuracy of the indirect approach. Side-by-side comparison of the two MS-based approaches for the first time further reveals unique advantages of the indirect approach, including better reproducibility and tolerance of interference. Moreover, the success of fishing out a potent ligand from a mixture of small-molecule fragments demonstrates great potential of the indirect LC/MS-based approach for constructing a robust screening platform against combinatorial libraries or natural product extracts. More importantly, by combining the results of MS-based analyses, enzymatic activity assay, competition experiments, and structural simulation, we discovered a new compound as a promising drug candidate targeting NDM-1.
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