Article (Scientific journals)
Biochemical and structural characterization of the subclass B1 metallo-β-lactamase VIM-4.
Lassaux, Patricia; Traoré, Daouda; Loisel, Elodie et al.
2011In Antimicrobial Agents and Chemotherapy
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Keywords :
Antibiotic resistance; carbapenemase; metallo-beta-lactamase
Abstract :
[en] The metallo-β-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 μM 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 Å. 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.
Disciplines :
Microbiology
Microbiology
Author, co-author :
Lassaux, Patricia ;  Université de Liège - ULiège > GIGA-R : Virologie - Immunologie
Traoré, Daouda
Loisel, Elodie
Favier, Alain
Docquier, Jean-Denis ;  Université de Liège - ULiège > Département des sciences de la vie > Centre d'ingénierie des protéines
Sohier, Jean ;  Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Laurent, Clémentine ;  Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Bebrone, Carine
Frère, Jean-Marie ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Ferrer, Jean-Louis
Galleni, Moreno ;  Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Language :
English
Title :
Biochemical and structural characterization of the subclass B1 metallo-β-lactamase VIM-4.
Publication date :
2011
Journal title :
Antimicrobial Agents and Chemotherapy
ISSN :
0066-4804
eISSN :
1098-6596
Publisher :
American Society for Microbiology (ASM), Washington, United States - District of Columbia
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 23 January 2015

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