Binding and processing of β-lactam antibiotics by the transpeptidase LdtMt2 from Mycobacterium tuberculosis
Steiner, Eva Maria LU
; Schneider, Gunter
and Schnell, Robert
(2017)
In FEBS Journal
284(5).
p.725-741
- Abstract
β-lactam antibiotics represent a novel direction in the chemotherapy of tuberculosis that brings the peptidoglycan layer of the complex mycobacterial cell wall in focus as a therapeutic target. Peptidoglycan stability in Mycobacterium tuberculosis, especially during infection, relies on the nonconventional peptide cross-links formed by l,d-transpeptidases. These enzymes are known to be inhibited by β-lactams, primarily carbapenems, leading to a stable covalent modification at the enzyme active site. A panel of 16 β-lactam antibiotics was characterized by inhibition kinetics, mass spectrometry, and x-ray crystallography to identify efficient compounds and study their action on the essential transpeptidase, LdtMt2. Members of... (More)
β-lactam antibiotics represent a novel direction in the chemotherapy of tuberculosis that brings the peptidoglycan layer of the complex mycobacterial cell wall in focus as a therapeutic target. Peptidoglycan stability in Mycobacterium tuberculosis, especially during infection, relies on the nonconventional peptide cross-links formed by l,d-transpeptidases. These enzymes are known to be inhibited by β-lactams, primarily carbapenems, leading to a stable covalent modification at the enzyme active site. A panel of 16 β-lactam antibiotics was characterized by inhibition kinetics, mass spectrometry, and x-ray crystallography to identify efficient compounds and study their action on the essential transpeptidase, LdtMt2. Members of the carbapenem class displayed fast binding kinetics, but faropenem, a penem type compound showed a three to four time higher rate in the adduct formation. In three cases, mass spectrometry indicated that carbapenems may undergo decarboxylation, while faropenem decomposition following the acylation step results in a small 87 Da β-OH-butyryl adduct bound at the catalytic cysteine residue. The crystal structure of LdtMt2 at 1.54 Å resolution with this fragment bound revealed that the protein adopts a closed conformation that shields the thioester bond from the solvent, which is in line with the high stability of this dead-end complex observed also in biochemical assays. Database: Structural data are available in Protein Data Bank under the accession numbers 5LB1 and 5LBG.
(Less)
- author
- Steiner, Eva Maria
LU
; Schneider, Gunter
and Schnell, Robert
- publishing date
- 2017-03-01
- type
- Contribution to journal
- publication status
- published
- keywords
- covalent adduct, faropenem, l,d-transpeptidase, protein structure, β-lactam antibiotic
- in
- FEBS Journal
- volume
- 284
- issue
- 5
- pages
- 725 - 741
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:85012048718
- pmid:28075068
- ISSN
- 1742-464X
- DOI
- 10.1111/febs.14010
- language
- English
- LU publication?
- no
- additional info
- Publisher Copyright: © 2017 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.
- id
- 0a271b72-86d9-4b14-9877-cf6338c69c69
- date added to LUP
- 2024-06-24 11:30:44
- date last changed
- 2024-06-25 03:04:13
@article{0a271b72-86d9-4b14-9877-cf6338c69c69,
abstract = {{<p>β-lactam antibiotics represent a novel direction in the chemotherapy of tuberculosis that brings the peptidoglycan layer of the complex mycobacterial cell wall in focus as a therapeutic target. Peptidoglycan stability in Mycobacterium tuberculosis, especially during infection, relies on the nonconventional peptide cross-links formed by l,d-transpeptidases. These enzymes are known to be inhibited by β-lactams, primarily carbapenems, leading to a stable covalent modification at the enzyme active site. A panel of 16 β-lactam antibiotics was characterized by inhibition kinetics, mass spectrometry, and x-ray crystallography to identify efficient compounds and study their action on the essential transpeptidase, Ldt<sub>Mt2</sub>. Members of the carbapenem class displayed fast binding kinetics, but faropenem, a penem type compound showed a three to four time higher rate in the adduct formation. In three cases, mass spectrometry indicated that carbapenems may undergo decarboxylation, while faropenem decomposition following the acylation step results in a small 87 Da β-OH-butyryl adduct bound at the catalytic cysteine residue. The crystal structure of Ldt<sub>Mt2</sub> at 1.54 Å resolution with this fragment bound revealed that the protein adopts a closed conformation that shields the thioester bond from the solvent, which is in line with the high stability of this dead-end complex observed also in biochemical assays. Database: Structural data are available in Protein Data Bank under the accession numbers 5LB1 and 5LBG.</p>}},
author = {{Steiner, Eva Maria and Schneider, Gunter and Schnell, Robert}},
issn = {{1742-464X}},
keywords = {{covalent adduct; faropenem; l,d-transpeptidase; protein structure; β-lactam antibiotic}},
language = {{eng}},
month = {{03}},
number = {{5}},
pages = {{725--741}},
publisher = {{Wiley-Blackwell}},
series = {{FEBS Journal}},
title = {{Binding and processing of β-lactam antibiotics by the transpeptidase Ldt<sub>Mt2</sub> from Mycobacterium tuberculosis}},
url = {{http://dx.doi.org/10.1111/febs.14010}},
doi = {{10.1111/febs.14010}},
volume = {{284}},
year = {{2017}},
}