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Nat Commun. 2017 Sep 14;8(1):538. doi: 10.1038/s41467-017-00601-9.

A general reaction mechanism for carbapenem hydrolysis by mononuclear and binuclear metallo-β-lactamases.

Author information

1
Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, S2002LRK, Rosario, Argentina.
2
Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, Montevideo, 11400, Uruguay.
3
Department of Chemistry and Biochemistry, Miami University, 651 E. High St., Oxford, OH, 45056, USA.
4
Instituto de Química Rosario (IQUIR, CONICET-UNR), Suipacha 531, Rosario, S2002LRK, Argentina.
5
Área Química General e Inorgánica, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, S2002LRK, Rosario, Argentina.
6
Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Cleveland, OH 44106, USA.
7
Departments of Medicine, Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
8
CARES, CWRU-VA Center for Antibiotic Resistance and Epidemiology, Cleveland, OH 44106, USA.
9
School of Cellular and Molecular Medicine, University of Bristol Biomedical Sciences Building, University Walk, Bristol, BS8 1TD, UK.
10
Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, S2002LRK, Rosario, Argentina. llarrull@ibr-conicet.gov.ar.
11
Area Biofísica, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, S2002LRK, Rosario, Argentina. llarrull@ibr-conicet.gov.ar.
12
Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET-UNR), Ocampo y Esmeralda, S2002LRK, Rosario, Argentina. vila@ibr-conicet.gov.ar.
13
CARES, CWRU-VA Center for Antibiotic Resistance and Epidemiology, Cleveland, OH 44106, USA. vila@ibr-conicet.gov.ar.
14
Area Biofísica, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, S2002LRK, Rosario, Argentina. vila@ibr-conicet.gov.ar.

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.Carbapenem-resistant bacteria pose a major health threat by expressing metallo-β-lactamases (MβLs), enzymes able to hydrolyse these life-saving drugs. Here the authors use biophysical and computational methods and show that different MβLs share the same reaction mechanism, suggesting new strategies for drug design.

PMID:
28912448
PMCID:
PMC5599593
DOI:
10.1038/s41467-017-00601-9
[Indexed for MEDLINE]
Free PMC Article

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