To probe the mechanism of inhibition of several previously-published metallo-β-lactamase (MBL) inhibitors for the clinically-important MBL Verona integron-encoded metallo-β-lactamase 2 (VIM-2), equilibrium dialyses with metal analyses, native state electrospray ionization mass spectrometry (ESI-MS), and UV–Vis spectrophotometry were utilized. The mechanisms of inhibition were analyzed for ethylenediaminetetraacetic acid (EDTA); dipicolinic acid (DPA) and DPA analogs 6-(1H-tetrazol-5-yl)picolinic acid (1T5PA) and 4-(3-aminophenyl)pyridine-2,6-dicarboxylic acid (3AP-DPA); thiol-containing compounds, 2,3-dimercaprol, thiorphan, captopril, and tiopronin; and 5-(pyridine-3-sulfonamido)-1,3-thiazole-4-carboxylic acid (ANT-431). UV–Vis spectroscopy and native-state ESI-MS results showed the formation of ternary complexes between VIM-2 and 1T5PA, ANT-431, thiorphan, captopril, and tiopronin, while a metal stripping mechanism was shown with VIM-2 and EDTA and DPA. The same approaches were used to show the formation of a ternary complex between New Delhi Metallo-β-lactamase (NDM-1) and ANT-431. The studies presented herein show that most of the inhibitors utilize a similar mechanism of inhibition as previously reported for NDM-1. These studies also demonstrate that native mass spectrometry can be used to probe the mechanism of inhibition at lower enzyme/inhibitor concentrations than has previously been achieved.

为了探讨几种先前发表的对临床上重要的MBL Verona整合子编码的金属β-内酰胺酶2（VIM-2）的金属β-内酰胺酶（MBL）抑制剂的抑制机理，采用金属分析的平衡透析液，原始状态使用电喷雾电离质谱（ESI-MS）和UV-Vis分光光度法。分析了乙二胺四乙酸（EDTA）的抑制机理；二吡啶甲酸（DPA）和DPA类似物6-（1H-四唑-5-基）吡啶甲酸（1T5PA）和4-（3-氨基苯基）吡啶-2,6-二羧酸（3AP-DPA）; 含硫醇的化合物，2，3-二巯基己醇，噻吩，卡托普利和硫普罗宁；和5-（吡啶-3-磺酰胺基）-1，3-噻唑-4-羧酸（ANT-431）。紫外可见光谱和原始状态的ESI-MS结果表明，VIM-2和1T5PA，ANT-431，硫醇，卡托普利和硫普罗宁，而VIM-2，EDTA和DPA则显示出金属剥离机理。使用相同的方法来显示新德里金属β-内酰胺酶（NDM-1）和ANT-431之间的三元复合物的形成。本文介绍的研究表明，大多数抑制剂利用与先前针对NDM-1报道的类似抑制机制。这些研究还表明，天然质谱可用于探测比以前已实现的酶/抑制剂浓度低的抑制机理。本文介绍的研究表明，大多数抑制剂利用与先前针对NDM-1报道的类似抑制机制。这些研究还表明，天然质谱可用于探测比以前已实现的酶/抑制剂浓度低的抑制机理。本文介绍的研究表明，大多数抑制剂利用与先前针对NDM-1报道的相似的抑制机制。这些研究还表明，天然质谱可用于探测比以前已实现的酶/抑制剂浓度低的抑制机理。