High-throughput screen with the l,d -transpeptidase Ldt Mt2 of Mycobacterium tuberculosis reveals novel classes of covalently reacting inhibitors
Disruption of bacterial cell wall biosynthesis in Mycobacterium tuberculosis is a promising target for treating tuberculosis. The l,d -transpeptidase Ldt Mt2 , which is responsible for the formation of 3 → 3 cross-links in the cell wall peptidoglycan, has been identified as essential for M. tubercul...
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Published in | Chemical science (Cambridge) Vol. 14; no. 26; pp. 7262 - 7278 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
05.07.2023
|
Online Access | Get full text |
ISSN | 2041-6520 2041-6539 |
DOI | 10.1039/D2SC06858C |
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Summary: | Disruption of bacterial cell wall biosynthesis in
Mycobacterium tuberculosis
is a promising target for treating tuberculosis. The
l,d
-transpeptidase Ldt
Mt2
, which is responsible for the formation of 3 → 3 cross-links in the cell wall peptidoglycan, has been identified as essential for
M. tuberculosis
virulence. We optimised a high-throughput assay for Ldt
Mt2
, and screened a targeted library of ∼10 000 electrophilic compounds. Potent inhibitor classes were identified, including established (
e.g.
, β-lactams) and unexplored covalently reacting electrophilic groups (
e.g.
, cyanamides). Protein-observed mass spectrometric studies reveal most classes to react covalently and irreversibly with the Ldt
Mt2
catalytic cysteine (Cys354). Crystallographic analyses of seven representative inhibitors reveal induced fit involving a loop enclosing the Ldt
Mt2
active site. Several of the identified compounds have a bactericidal effect on
M. tuberculosis
within macrophages, one with an MIC
50
value of ∼1 μM. The results provide leads for the development of new covalently reaction inhibitors of Ldt
Mt2
and other nucleophilic cysteine enzymes. |
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ISSN: | 2041-6520 2041-6539 |
DOI: | 10.1039/D2SC06858C |