A Fluorescence‐based assay for screening β‐lactams targeting the Mycobacterium tuberculosis transpeptidase LdtMt2
| Autor: | Mariska de Munnik, Christopher J. Schofield, Christopher T. Lohans, Corentin Bon, Jürgen Brem, Gareth W. Langley |
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| Přispěvatelé: | Chemistry Research Laboratory [Oxford, UK], University of Oxford [Oxford], Queen's University [Kingston, Canada], This project was co-funded by the Tres Cantos Open Lab Foundation (ProjectTC241). We thank the Wellcome Trust and the Medical Research Council (MRC) for supporting our work on antibiotics. |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Fluorophore
Tuberculosis medicine.drug_class [SDV]Life Sciences [q-bio] Antibiotics 010402 general chemistry 01 natural sciences Biochemistry Subclass antibiotics Mycobacterium tuberculosis chemistry.chemical_compound Biosynthesis fluorescent probes inhibitors medicine Potency [CHIM]Chemical Sciences Molecular Biology chemistry.chemical_classification biology 010405 organic chemistry beta-lactams Organic Chemistry medicine.disease biology.organism_classification 0104 chemical sciences 3. Good health Enzyme chemistry tuberculosis Molecular Medicine |
| Zdroj: | ChemBioChem ChemBioChem, Wiley-VCH Verlag, 2020, 21 (3), pp.368-372. ⟨10.1002/cbic.201900379⟩ |
| ISSN: | 1439-7633 1439-4227 |
| Popis: | International audience; 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. |
| Databáze: | OpenAIRE |
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