Mapping the determinants of catalysis and substrate specificity of the antibiotic resistance enzyme CTX-M β-lactamase
Abstract
Abstract CTX-M β-lactamases are prevalent antibiotic resistance enzymes and are notable for their ability to rapidly hydrolyze the extended-spectrum cephalosporin, cefotaxime. We hypothesized that the active site sequence requirements of CTX-M-mediated hydrolysis differ between classes of β-lactam antibiotics. Accordingly, we use codon randomization, antibiotic selection, and deep sequencing to determine the CTX-M active-site residues required for hydrolysis of cefotaxime and the penicillin, ampicillin. The study reveals positions required for hydrolysis of all β-lactams, as well as residues controlling substrate specificity. Further, CTX-M enzymes poorly hydrolyze the extended-spectrum cephalosporin, ceftazidime. We further show that the sequence requirements for ceftazidime hydrolysis follow those of cefotaxime, with the exception that key active-site omega loop residues are not required, and may be detrimental, for ceftazidime hydrolysis. These results provide insights into cephalosporin hydrolysis and demonstrate that changes to the active-site omega loop are likely required for the evolution of CTX-M-mediated ceftazidime resistance.
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC); National Institutes of Health (NIH); Welch Foundation
- OSTI Identifier:
- 1909043
- Alternate Identifier(s):
- OSTI ID: 1972190
- Grant/Contract Number:
- AC02-05CH11231; AI32956; Q1279; T32 GM120011; P30 GM124169-01
- Resource Type:
- Published Article
- Journal Name:
- Communications Biology
- Additional Journal Information:
- Journal Name: Communications Biology Journal Volume: 6 Journal Issue: 1; Journal ID: ISSN 2399-3642
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; antibiotics; combinatorial libraries; enzyme mechanisms; hydrolases; x-ray crystallography
Citation Formats
Judge, Allison, Hu, Liya, Sankaran, Banumathi, Van Riper, Justin, Venkataram Prasad, B. V., and Palzkill, Timothy. Mapping the determinants of catalysis and substrate specificity of the antibiotic resistance enzyme CTX-M β-lactamase. United Kingdom: N. p., 2023.
Web. doi:10.1038/s42003-023-04422-z.
Judge, Allison, Hu, Liya, Sankaran, Banumathi, Van Riper, Justin, Venkataram Prasad, B. V., & Palzkill, Timothy. Mapping the determinants of catalysis and substrate specificity of the antibiotic resistance enzyme CTX-M β-lactamase. United Kingdom. https://doi.org/10.1038/s42003-023-04422-z
Judge, Allison, Hu, Liya, Sankaran, Banumathi, Van Riper, Justin, Venkataram Prasad, B. V., and Palzkill, Timothy. Thu .
"Mapping the determinants of catalysis and substrate specificity of the antibiotic resistance enzyme CTX-M β-lactamase". United Kingdom. https://doi.org/10.1038/s42003-023-04422-z.
@article{osti_1909043,
title = {Mapping the determinants of catalysis and substrate specificity of the antibiotic resistance enzyme CTX-M β-lactamase},
author = {Judge, Allison and Hu, Liya and Sankaran, Banumathi and Van Riper, Justin and Venkataram Prasad, B. V. and Palzkill, Timothy},
abstractNote = {Abstract CTX-M β-lactamases are prevalent antibiotic resistance enzymes and are notable for their ability to rapidly hydrolyze the extended-spectrum cephalosporin, cefotaxime. We hypothesized that the active site sequence requirements of CTX-M-mediated hydrolysis differ between classes of β-lactam antibiotics. Accordingly, we use codon randomization, antibiotic selection, and deep sequencing to determine the CTX-M active-site residues required for hydrolysis of cefotaxime and the penicillin, ampicillin. The study reveals positions required for hydrolysis of all β-lactams, as well as residues controlling substrate specificity. Further, CTX-M enzymes poorly hydrolyze the extended-spectrum cephalosporin, ceftazidime. We further show that the sequence requirements for ceftazidime hydrolysis follow those of cefotaxime, with the exception that key active-site omega loop residues are not required, and may be detrimental, for ceftazidime hydrolysis. These results provide insights into cephalosporin hydrolysis and demonstrate that changes to the active-site omega loop are likely required for the evolution of CTX-M-mediated ceftazidime resistance.},
doi = {10.1038/s42003-023-04422-z},
journal = {Communications Biology},
number = 1,
volume = 6,
place = {United Kingdom},
year = {Thu Jan 12 00:00:00 EST 2023},
month = {Thu Jan 12 00:00:00 EST 2023}
}
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