TY - JOUR
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)
C2 - 28912448
AN - SCOPUS:85029603076
VL - 8
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
M1 - 538
ER -