The mechanism of NDM-1-catalyzed carbapenem hydrolysis is distinct from that of penicillin or cephalosporin hydrolysis

Nat Commun. 2017 Dec 21;8(1):2242. doi: 10.1038/s41467-017-02339-w.

Abstract

New Delhi metallo-β-lactamases (NDMs), the recent additions to metallo-β-lactamases (MBLs), pose a serious public health threat due to its highly efficient hydrolysis of β-lactam antibiotics and rapid worldwide dissemination. The MBL-hydrolyzing mechanism for carbapenems is less studied than that of penicillins and cephalosporins. Here, we report crystal structures of NDM-1 in complex with hydrolyzed imipenem and meropenem, at resolutions of 1.80-2.32 Å, together with NMR spectra monitoring meropenem hydrolysis. Three enzyme-intermediate/product derivatives, EI1, EI2, and EP, are trapped in these crystals. Our structural data reveal double-bond tautomerization from Δ2 to Δ1, absence of a bridging water molecule and an exclusive β-diastereomeric product, all suggesting that the hydrolytic intermediates are protonated by a bulky water molecule incoming from the β-face. These results strongly suggest a distinct mechanism of NDM-1-catalyzed carbapenem hydrolysis from that of penicillin or cephalosporin hydrolysis, which may provide a novel rationale for design of mechanism-based inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbapenems / metabolism*
  • Cephalosporins / metabolism*
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli
  • Hydrolysis
  • Imipenem / metabolism
  • Magnetic Resonance Spectroscopy
  • Meropenem
  • Models, Molecular
  • Penicillins / metabolism*
  • Thienamycins / metabolism
  • beta-Lactamases / metabolism*

Substances

  • Carbapenems
  • Cephalosporins
  • Penicillins
  • Thienamycins
  • Imipenem
  • beta-Lactamases
  • beta-lactamase NDM-1
  • Meropenem