title: Identification of inhibitors of the antibiotic-resistance target New Delhi metallo-β-lactamase 1 by both nanoelectrospray ionization mass spectrometry and ultrafiltration liquid chromatography/mass spectrometry approaches.
abstract: 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-β-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.
authors.affiliation: College of Life Sciences and Tianjin State Laboratory of Protein Science, Nankai University, Tianjin, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.forename: Lixin
authors.initials: L
authors.lastname: Li
authors.forename: Shuai
authors.initials: S
authors.lastname: Chen
authors.forename: Yintong
authors.initials: Y
authors.lastname: Xu
authors.forename: Qiang
authors.initials: Q
authors.lastname: Xia
authors.forename: Yu
authors.initials: Y
authors.lastname: Guo
authors.forename: Xiang
authors.initials: X
authors.lastname: Liu
authors.forename: Yanting
authors.initials: Y
authors.lastname: Tang
authors.forename: Tanjie
authors.initials: T
authors.lastname: Zhang
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Cheng
authors.initials: C
authors.lastname: Yang
authors.forename: Wenqing
authors.initials: W
authors.lastname: Shui
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002853:Chromatography, Liquid / Q000379:methods*; D004352:Drug Resistance, Microbial; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D036103:Nanotechnology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014462:Ultrafiltration / Q000379:methods*; D001618:beta-Lactamases / Q000032:analysis*
chemical_list: D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D001618:beta-Lactamases; C552864:beta-lactamase NDM-1