[en] Among class B beta-lactamases, the subclass B2 CphA enzyme is characterized by a unique specificity profile. CphA efficiently hydrolyzes only carbapenems. In this work, we generated site-directed mutants that possess a strongly broadened activity spectrum when compared with the WT CphA. Strikingly, the N116H/N220G double mutant exhibits a substrate profile close to that observed for the broad spectrum subclass B1 enzymes. The double mutant is significantly activated by the binding of a second zinc ion under conditions where the WT enzyme is non-competitively inhibited by the same ion.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Bebrone, Carine ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Anne, Christine ; Université de Liège - ULiège > Centre d'ingénierie des protéines
De Vriendt, K.
Devreese, B.
Rossolini, G. M.
Van Beeumen, Jos
Frère, Jean-Marie ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Galleni, Moreno ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Language :
English
Title :
Dramatic broadening of the substrate profile of the Aeromonas hydrophila CphA metallo-beta-lactamase by site-directed mutagenesis
Publication date :
05 August 2005
Journal title :
Journal of Biological Chemistry
ISSN :
0021-9258
eISSN :
1083-351X
Publisher :
Amer Soc Biochemistry Molecular Biology Inc, Bethesda, United States - Maryland
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