A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase LdtMt2
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Authors
de Munnik, Mariska
Lohans, Christopher T.
Langley, Gareth W.
Bon, Corentin
Brem, Jürgen
Schofield, Christopher J.
Date
2019
Type
preprint
Language
en
Keyword
Antibiotics , Beta-Lactams , Fluorescent Probes , Inhibitors , Tuberculosis
Alternative Title
Abstract
Mycobacterium tuberculosis l,d‐transpeptidases (Ldts), which are involved in cell‐wall biosynthesis, have emerged as promising targets for the treatment of tuberculosis. However, an efficient method for testing inhibition of these enzymes is not currently available. We present a fluorescence‐based assay for LdtMt2, which is suitable for high‐throughput screening. Two fluorogenic probes were identified that release a fluorophore upon reaction with LdtMt2, thus making it possible to assess the availability of the catalytic site in the presence of inhibitors. The assay was applied to a panel of β‐lactam antibiotics and related inhibitors; the results validate observations that the (carba)penem subclass of β‐lactams are more potent Ldt inhibitors than other β‐lactam classes, though unexpected variations in potency were observed. The method will enable systematic structure–activity relationship studies on Ldts, thereby facilitating the identification of new antibiotics active against M. tuberculosis.
Description
Citation
Publisher
Wiley
License
This is the peer reviewed version of the following article: de Munnik et a. A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase LdtMt2, which has been published in final form at https://doi.org/10.1002/cbic.201900379. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.