Inhibition of the Broad Spectrum Nonmetallocarbapenamase of Class A (NMC-A) ␤-Lactamase from Enterobacter cloacae by Monocyclic ␤-Lactams* □ S - Archive ouverte HAL Access content directly
Journal Articles Journal of Biological Chemistry Year : 1999

Inhibition of the Broad Spectrum Nonmetallocarbapenamase of Class A (NMC-A) ␤-Lactamase from Enterobacter cloacae by Monocyclic ␤-Lactams* □ S

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1
Lakshmi P Kotra
  • Function : Author
John Bellettiniʈ
  • Function : Author
Alexey Bulychev
  • Function : Author
Michael O'Brien
  • Function : Author
Marvin J Millerʈ
  • Function : Author
Shahriar Mobashery
  • Function : Author
Jean-Pierre Samama
  • Function : Author
  • PersonId : 1081525

Abstract

␤-Lactamases hydrolyze ␤-lactam antibiotics, a reaction that destroys their antibacterial activity. These enzymes , of which four classes are known, are the primary cause of resistance to ␤-lactam antibiotics. The class A ␤-lactamases form the largest group. A novel class A ␤-lactamase, named the nonmetallocarbapenamase of class A (NMC-A) ␤-lactamase, has been discovered recently that has a broad substrate profile that included carbapenem antibiotics. This is a serious development, since carbapenems have been relatively immune to the action of these resistance enzymes. Inhibitors for this enzyme are sought. We describe herein that a type of monobactam molecule of our design inactivates the NMC-A ␤-lactamase rapidly, efficiently, and irreversibly. The mechanism of inactivation was investigated by solving the x-ray structure of the inhibited NMC-A enzyme to 1.95 Å resolution. The structure shed light on the nature of the fragmentation of the inhibitor on enzyme acylation and indicated that there are two acyl-enzyme species that account for enzyme inhibition. Each of these inhibited enzyme species is trapped in a distinct local energy minimum that does not predispose the inhibitor species for deacylation, accounting for the irreversible mode of enzyme inhibition. Molecular dynamics simulations provided evidence in favor of a dynamic motion for the acyl-enzyme species, which samples a considerable conformational space prior to the entrapment of the two stable acyl-enzyme species in the local energy minima. A discussion of the likelihood of such dynamic motion for turnover of substrates during the normal catalytic processes of the enzyme is presented.
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hal-03004384 , version 1 (20-11-2020)

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  • HAL Id : hal-03004384 , version 1

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Lionel Mourey, Lakshmi P Kotra, John Bellettiniʈ, Alexey Bulychev, Michael O'Brien, et al.. Inhibition of the Broad Spectrum Nonmetallocarbapenamase of Class A (NMC-A) ␤-Lactamase from Enterobacter cloacae by Monocyclic ␤-Lactams* □ S. Journal of Biological Chemistry, 1999, 274, pp.25260 - 25265. ⟨hal-03004384⟩
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