Kinetic Study of Laboratory Mutants of NDM-1 Metallo-beta-Lactamase and the Importance of an Isoleucine at Position 35.

[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 :

Issue :

Pages :

2366-72

Peer reviewed :

Peer Reviewed verified by ORBi

Commentary :

Available on ORBi :

since 04 April 2016

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