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Real‐Time Monitoring of New Delhi Metallo‐β‐Lactamase Activity in Living Bacterial Cells by 1 H NMR Spectroscopy
Author(s) -
Ma Junhe,
McLeod Sarah,
MacCormack Kathleen,
Sriram Shubha,
Gao Ning,
Breeze Alexander L.,
Hu Jun
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201308636
Subject(s) - escherichia coli , meropenem , ethylenediaminetetraacetic acid , in vivo , chemistry , spermine , nuclear magnetic resonance spectroscopy , in vitro , biochemistry , bacteria , microbiology and biotechnology , hydrolysis , antibiotics , stereochemistry , biology , antibiotic resistance , enzyme , organic chemistry , chelation , genetics , gene
Abstract Disconnections between in vitro responses and those observed in whole cells confound many attempts to design drugs in areas of serious medical need. A method based on 1D 1 H 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‐β‐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 in vivo. NDM‐1 activity in cells was also shown to be inhibited by spermine, a porin inhibitor, although in an in vitro 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.