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

Mariá Natalia Lisa; Antonela R. Palacios; Mahesh Aitha; Mariano M. González; DIego M. Moreno; Michael W. Crowder; Robert A. Bonomo; James Spencer; David L. Tierney; Leticia I. Llarrull; Alejandro J. Vila

doi:10.1038/s41467-017-00601-9

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

Mariá Natalia Lisa, Antonela R. Palacios, Mahesh Aitha, Mariano M. González, DIego M. Moreno, Michael W. Crowder, Robert A. Bonomo, James Spencer, David L. Tierney, Leticia I. Llarrull, Alejandro J. Vila^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

51 Citations (Scopus)

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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.

Original language	English
Article number	538
Journal	Nature Communications
Volume	8
DOIs	https://doi.org/10.1038/s41467-017-00601-9
Publication status	Published - 14 Sep 2017

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Lisa, Mariá Natalia ; Palacios, Antonela R. ; Aitha, Mahesh ; González, Mariano M. ; Moreno, DIego M. ; Crowder, Michael W. ; Bonomo, Robert A. ; Spencer, James ; Tierney, David L. ; Llarrull, Leticia I. ; Vila, Alejandro J. / A general reaction mechanism for carbapenem hydrolysis by mononuclear and binuclear metallo-β-lactamases. In: Nature Communications. 2017 ; Vol. 8.

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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.",

author = "Lisa, {Mari{\'a} Natalia} and Palacios, {Antonela R.} and Mahesh Aitha and Gonz{\'a}lez, {Mariano M.} and Moreno, {DIego M.} and Crowder, {Michael W.} and Bonomo, {Robert A.} and James Spencer and Tierney, {David L.} and Llarrull, {Leticia I.} and Vila, {Alejandro J.}",

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A general reaction mechanism for carbapenem hydrolysis by mononuclear and binuclear metallo-β-lactamases. / Lisa, Mariá Natalia; Palacios, Antonela R.; Aitha, Mahesh; González, Mariano M.; Moreno, DIego M.; Crowder, Michael W.; Bonomo, Robert A.; Spencer, James; Tierney, David L.; Llarrull, Leticia I.; Vila, Alejandro J.

In: Nature Communications, Vol. 8, 538, 14.09.2017.

Research output: Contribution to journal › Article (Academic Journal) › peer-review

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AB - 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.

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DO - 10.1038/s41467-017-00601-9

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