[en] Two laboratory mutants of NDM-1 were generated by replacing the isoleucine at position 35 with threonine and serine residues: the NDM-1(I35T)and NDM-1(I35S)enzymes. These mutants were well characterized, and their kinetic parameters were compared with those of the NDM-1 wild type. Thekcat,Km, andkcat/Kmvalues calculated for the two mutants were slightly different from those of the wild-type enzyme. Interestingly, thekcat/Kmof NDM-1(I35S)for loracarbef was about 14-fold higher than that of NDM-1. Far-UV circular dichroism (CD) spectra of NDM-1 and NDM-1(I35T)and NDM-1(I35S)enzymes suggest local structural rearrangements in the secondary structure with a marked reduction of alpha-helix content in the mutants.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Marcoccia, Francesca
Bottoni, Carlo
Sabatini, Alessia
Colapietro, Martina
Mercuri, Paola ; Université de Liège > Département des sciences de la vie > Macromolécules biologiques
Galleni, Moreno ; Université de Liège > Département des sciences de la vie > Macromolécules biologiques
Kerff, Frédéric ; Université de Liège > Département des sciences de la vie > Centre d'ingénierie des protéines
Matagne, André ; Université de Liège > Département des sciences de la vie > Enzymologie et repliement des protéines
Celenza, Giuseppe
Amicosante, Gianfranco
Perilli, Mariagrazia
Language :
English
Title :
Kinetic Study of Laboratory Mutants of NDM-1 Metallo-beta-Lactamase and the Importance of an Isoleucine at Position 35.
Publication date :
2016
Journal title :
Antimicrobial Agents and Chemotherapy
ISSN :
0066-4804
eISSN :
1098-6596
Publisher :
American Society for Microbiology, Washington, United States - District of Columbia
Volume :
60
Issue :
4
Pages :
2366-72
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright (c) 2016, American Society for Microbiology. All Rights Reserved.
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