Biochemical and structural characterization of the subclass B1 metallo-beta-lactamase VIM-4

Patricia Lassaux, Daouda A K Traore, Elodie Loisel, Jean -Denis Docquier, Jean Sebasiten Sohier, Clementine Laurent, Carine Bebrone, Jean -Marie Frere, Jean -Luc Ferrer, Moreno Galleni

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Abstract

The metallo-beta-lactamase VIM-4, mainly found in Pseudomonas aeruginosa or Acinetobacter baumannii, was produced in Escherichia coli and characterized by biochemical and X-ray techniques. A detailed kinetic study performed in the presence of Zn2+ at concentrations ranging from 0.4 to 100 uM showed that VIM-4 exhibits a kinetic profile similar to the profiles of VIM-2 and VIM-1. However, VIM-4 is more active than VIM-1 against benzylpenicillin, cephalothin, nitrocefin, and imipenem and is less active than VIM-2 against ampicillin and meropenem. The crystal structure of the dizinc form of VIM-4 was solved at 1.9 ??. The sole difference between VIM-4 and VIM-1 is found at residue 228, which is Ser in VIM-1 and Arg in VIM-4. This substitution has a major impact on the VIM-4 catalytic efficiency compared to that of VIM-1. In contrast, the differences between VIM-2 and VIM-4 seem to be due to a different position of the flapping loop and two substitutions in loop 2. Study of the thermal stability and the activity of the holo- and apo-VIM-4 enzymes revealed that Zn2+ ions have a pronounced stabilizing effect on the enzyme and are necessary for preserving the structure.
Original languageEnglish
Pages (from-to)1248 - 1255
Number of pages8
JournalAntimicrobial Agents and Chemotherapy
Volume55
Issue number3
DOIs
Publication statusPublished - 2011
Externally publishedYes

Cite this

Lassaux, P., Traore, D. A. K., Loisel, E., Docquier, J. -D., Sohier, J. S., Laurent, C., Bebrone, C., Frere, J. -M., Ferrer, J. -L., & Galleni, M. (2011). Biochemical and structural characterization of the subclass B1 metallo-beta-lactamase VIM-4. Antimicrobial Agents and Chemotherapy, 55(3), 1248 - 1255. https://doi.org/10.1128/AAC.01486-09