Real-Time Monitoring of New Delhi Metallo-β-Lactamase Activity in Living Bacterial Cells by 1H NMR Spectroscopy
Real-Time Monitoring of New Delhi Metallo-β-Lactamase Activity in Living Bacterial Cells by 1H NMR Spectroscopy
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DOI：
10.1002/anie.201308636
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发表时间：
2014-02-17
影响因子：
16.6
通讯作者：
Hu, Jun
中科院分区：
文献类型：
--
作者：
Ma, Junhe;McLeod, Sarah;Hu, Jun
体外反应和在整个细胞中观察到的反应之间的脱节，混淆了在严重医疗需求领域设计药物的许多尝试。报道了一种基于1DH-1核磁共振波谱的方法，该方法能够监测碳青霉烯类抗生素美罗培南在表达新德里金属内酰胺酶亚类1(NDM-1)的大肠杆菌细胞中的水解分解，这是一种新的抗生素耐药威胁。基于细胞的核磁共振研究表明，已知的两种NDM-1抑制剂L-卡托普利和乙二胺四乙酸在体内都能抑制美罗培南的水解。尽管在体外实验中，精胺对分离的NDM-1蛋白的活性影响很小，但细胞内的NDM-1活性也被孔蛋白抑制剂精胺抑制。这一新方法可能对监测其他复杂生物基质和包括哺乳动物细胞在内的全细胞环境中涉及可扩散代谢物的反应具有普遍实用价值。
Disconnections between invitro responses and those observed in whole cells confound many attempts to design drugs in areas of serious medical need. A method based on 1D H-1 NMR spectroscopy is reported that affords the ability to monitor the hydrolytic decomposition of the carbapenem antibiotic meropenem inside Escherichia coli cells expressing New Delhi metallo--lactamase subclass 1 (NDM-1), an emerging antibiotic-resistance threat. Cell-based NMR studies demonstrated that two known NDM-1 inhibitors, L-captopril and ethylenediaminetetraacetic acid (EDTA), inhibit the hydrolysis of meropenem invivo. NDM-1 activity in cells was also shown to be inhibited by spermine, a porin inhibitor, although in an invitro assay, the influence of spermine on the activity of isolated NDM-1 protein is minimal. This new approach may have generic utility for monitoring reactions involving diffusible metabolites in other complex biological matrices and whole-cell settings, including mammalian cells.