Article (Scientific journals)
On functional and structural heterogeneity of VIM-type metallo-beta-lactamases
Docquier, Jean-Denis; Lamotte-Brasseur, Josette; Galleni, Moreno et al.
2003In Journal of Antimicrobial Chemotherapy, 51 (2), p. 257-266
Peer Reviewed verified by ORBi
 

Files


Full Text
Docquier_2003.pdf
Publisher postprint (2.82 MB)
Request a copy

All documents in ORBi are protected by a user license.

Send to



Details



Abstract :
[en] The VIM metallo-beta-lactamases are emerging resistance determinants, encoded by mobile genetic elements, that have recently been detected in multidrug-resistant nosocomial isolates of Pseudomonas aeruginosa and other Gram-negative pathogens. In this work a T7-based expression system for overproduction of the VIM-2 enzyme by Escherichia coli was developed, which yielded similar to80 mg of protein per litre of culture. The enzyme was mostly released into the medium, from which it was recovered at >99% purity by an initial ammonium sulphate precipitation followed by two chromatography steps, with almost 80% efficiency. Determination of kinetic parameters of VIM-2 under the same experimental conditions previously used for VIM-1 (the first VIM-type enzyme detected in clinical isolates, which is 93% identical to VIM-2) revealed significant differences in K-m values and/or turnover rates with several substrates, including penicillins, cephalosporins and carbapenems. Compared with VIM-1, VIM-2 is more susceptible to inactivation by chelators, indicating that the zinc ions of the latter are probably more loosely bound. These data indicated that at least some of the amino acid differences between the two proteins have functional significance. Molecular modelling of the two enzymes identified some amino acid substitutions, including those at positions 223, 224 and 228 (in the BBL numbering), that could be relevant to the changes in catalytic behaviour.
Disciplines :
Pharmacy, pharmacology & toxicology
Microbiology
Immunology & infectious disease
Author, co-author :
Docquier, Jean-Denis ;  Université de Liège - ULiège > Département des sciences de la vie > Centre d'ingénierie des protéines
Lamotte-Brasseur, Josette 
Galleni, Moreno ;  Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Amicosante, G.
Frère, Jean-Marie ;  Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Rossolini, G. M.
Language :
English
Title :
On functional and structural heterogeneity of VIM-type metallo-beta-lactamases
Publication date :
February 2003
Journal title :
Journal of Antimicrobial Chemotherapy
ISSN :
0305-7453
eISSN :
1460-2091
Publisher :
Oxford Univ Press, Oxford, United Kingdom
Volume :
51
Issue :
2
Pages :
257-266
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 24 November 2015

Statistics


Number of views
52 (5 by ULiège)
Number of downloads
3 (3 by ULiège)

Scopus citations®
 
145
Scopus citations®
without self-citations
101
OpenCitations
 
134

Bibliography


Similar publications



Contact ORBi