Abstract

Carbapenem-resistant Enterobacteriaceae threaten human health, since carbapenems are last resort drugs for infections by such organisms. Metallo-β-lactamases (MβLs) are the main mechanism of resistance against carbapenems. Clinically approved inhibitors of MBLs are currently unavailable as design has been limited by the incomplete knowledge of their mechanism. Here, we report a biochemical and biophysical study of carbapenem hydrolysis by the B1 enzymes NDM-1 and BcII in the bi-Zn(II) form, the mono-Zn(II) B2 Sfh-I and the mono-Zn(II) B3 GOB-18. These MβLs hydrolyse carbapenems via a similar mechanism, with accumulation of the same anionic intermediates. We characterize the Michaelis complex formed by mono-Zn(II) enzymes, and we identify all intermediate species, enabling us to propose a chemical mechanism for mono and binuclear MβLs. This common mechanism open avenues for rationally designed inhibitors of all MβLs, notwithstanding the profound differences between these enzymes’ active site structure, β-lactam specificity and metal content.

Details

Title
A general reaction mechanism for carbapenem hydrolysis by mononuclear and binuclear metallo-β-lactamases
Author
María-Natalia, Lisa 1   VIAFID ORCID Logo  ; Palacios, Antonela R 2 ; Aitha, Mahesh 3 ; González, Mariano M 2 ; Moreno, Diego M 4 ; Crowder, Michael W 3 ; Bonomo, Robert A 5 ; Spencer, James 6 ; Tierney, David L 3 ; Llarrull, Leticia I 7 ; Vila, Alejandro J 8 

1  Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, Rosario, Argentina; Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, Montevideo, Uruguay 
2  Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, Rosario, Argentina 
3  Department of Chemistry and Biochemistry, Miami University, Oxford, OH, USA 
4  Instituto de Química Rosario (IQUIR, CONICET-UNR), Rosario, Argentina; Área Química General e Inorgánica, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina 
5  Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Cleveland, USA; Departments of Medicine, Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, USA; CARES, CWRU-VA Center for Antibiotic Resistance and Epidemiology, Cleveland, USA 
6  School of Cellular and Molecular Medicine, University of Bristol Biomedical Sciences Building, University Walk, Bristol, UK 
7  Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, Rosario, Argentina; Area Biofísica, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina 
8  Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, Rosario, Argentina; CARES, CWRU-VA Center for Antibiotic Resistance and Epidemiology, Cleveland, USA; Area Biofísica, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina 
Pages
1-11
Publication year
2017
Publication date
Sep 2017
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-017-00601-9
ProQuest document ID
1938826320
Copyright
© 2017. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.