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

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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 Sept 2017

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@article{786de1a5846b40959742fdeeb17f03a4,

title = "A general reaction mechanism for carbapenem hydrolysis by mononuclear and binuclear metallo-β-lactamases",

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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T1 - A general reaction mechanism for carbapenem hydrolysis by mononuclear and binuclear metallo-β-lactamases

AU - Lisa, Mariá Natalia

AU - Palacios, Antonela R.

AU - Aitha, Mahesh

AU - González, Mariano M.

AU - Moreno, DIego M.

AU - Crowder, Michael W.

AU - Bonomo, Robert A.

AU - Spencer, James

AU - Tierney, David L.

AU - Llarrull, Leticia I.

AU - Vila, Alejandro J.

PY - 2017/9/14

Y1 - 2017/9/14

N2 - 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.

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.

U2 - 10.1038/s41467-017-00601-9

DO - 10.1038/s41467-017-00601-9

M3 - Article (Academic Journal)

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AN - SCOPUS:85029603076

SN - 2041-1723

VL - 8

JO - Nature Communications

JF - Nature Communications

M1 - 538

ER -

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

Abstract

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