Molecular dynamics at the root of expansion of function in the M69L inhibitor-resistant TEM β-lactamase from Escherichia coli

Samy O. Meroueh, Pierre Roblin, Dasantila Golemi, Laurent Maveyraud, Sergei B. Vakulenko, Yun Zhang, Jean Pierre Samama, Shahriar Mobashery

Research output: Contribution to journalArticle

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Abstract

Clavulanate, an inhibitor for β-lactamases, was the very first inhibitor for an antibiotic resistance enzyme that found clinical utility in 1985. The clinical use of clavulanate and that of sulbactam and tazobactam, which were introduced to the clinic subsequently, has facilitated evolution of a set of β-lactamases that not only retain their original function as resistance enzymes but also are refractory to inhibition by the inhibitors. This article characterizes the properties of the clinically identified M69L mutant variant of the TEM-1 β-lactamase from Escherichia coli, an inhibitor-resistant β-lactamase, and compares it to the wild-type enzyme. The enzyme is as active as the wild-type in turnover of typical β-lactam antibiotics. Furthermore, many of the parameters for interactions of the inhibitors with the mutant enzyme are largely unaffected. The significant effect of the inhibitor-resistant trait was a relatively modest elevation of the dissociation constant for the formation of the pre-acylation complex. The high-resolution x-ray crystal structure for the M69L mutant variant revealed essentially no alteration of the three-dimensional structure, both for the protein backbone and for the positions of the side chains of the amino acids. It was surmised that the difference in the two enzymes must reside with the dynamic motions of the two proteins. Molecular dynamics simulations of the mutant and wild-type proteins were carried out for 2 ns each. Dynamic cross-correlated maps revealed the collective motions of the two proteins to be very similar, yet the two proteins did not behave identically. Differences in behavior of the two proteins existed in the regions between residues 145-179 and 155-162. Additional calculations revealed that kinetic effects measured experimentally for the dissociation constant for the pre-acylation complex could be mostly attributed to the electrostatic and van der Waals components of the binding free energy. The effects of the mutation on the behavior of the β-lactamase were subtle, including the differences in the measured dissociation constants that account for the inhibitor-resistant trait. It would appear that nature has selected for incorporation of the most benign alteration in the structure of the wild-type TEM-1 β-lactamase that is sufficient to give the inhibitor-resistant trait.

Original languageEnglish (US)
Pages (from-to)9422-9430
Number of pages9
JournalJournal of the American Chemical Society
Volume124
Issue number32
DOIs
StatePublished - Aug 14 2002

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Molecular Dynamics Simulation
Escherichia coli
Molecular dynamics
Enzymes
Transmission electron microscopy
Proteins
Acylation
Clavulanic Acid
Antibiotics
Anti-Bacterial Agents
Enzyme inhibition
Sulbactam
Lactams
Mutant Proteins
Microbial Drug Resistance
Static Electricity
Refractory materials
Free energy
Amino acids
Electrostatics

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Molecular dynamics at the root of expansion of function in the M69L inhibitor-resistant TEM β-lactamase from Escherichia coli. / Meroueh, Samy O.; Roblin, Pierre; Golemi, Dasantila; Maveyraud, Laurent; Vakulenko, Sergei B.; Zhang, Yun; Samama, Jean Pierre; Mobashery, Shahriar.

In: Journal of the American Chemical Society, Vol. 124, No. 32, 14.08.2002, p. 9422-9430.

Research output: Contribution to journalArticle

Meroueh, Samy O. ; Roblin, Pierre ; Golemi, Dasantila ; Maveyraud, Laurent ; Vakulenko, Sergei B. ; Zhang, Yun ; Samama, Jean Pierre ; Mobashery, Shahriar. / Molecular dynamics at the root of expansion of function in the M69L inhibitor-resistant TEM β-lactamase from Escherichia coli. In: Journal of the American Chemical Society. 2002 ; Vol. 124, No. 32. pp. 9422-9430.
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AU - Maveyraud, Laurent

AU - Vakulenko, Sergei B.

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AU - Samama, Jean Pierre

AU - Mobashery, Shahriar

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