Dramatic broadening of the substrate profile of the Aeromonas hydrophila CphA metallo-β-lactamase by site-directed mutagenesis

Carine Bebrone, Christine Anne, Kris De Vriendt, Bart Devreese, Gian Maria Rossolini, Jozef Van Beeumen, Jean Marie Frère, Moreno Galleni

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Among class B β3-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.

Original languageEnglish
Pages (from-to)28195-28202
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number31
DOIs
Publication statusPublished - Aug 5 2005

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Aeromonas hydrophila
Mutagenesis
Site-Directed Mutagenesis
Substrates
Enzymes
Ions
Carbapenems
Zinc

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bebrone, C., Anne, C., De Vriendt, K., Devreese, B., Rossolini, G. M., Van Beeumen, J., ... Galleni, M. (2005). Dramatic broadening of the substrate profile of the Aeromonas hydrophila CphA metallo-β-lactamase by site-directed mutagenesis. Journal of Biological Chemistry, 280(31), 28195-28202. https://doi.org/10.1074/jbc.M414052200

Dramatic broadening of the substrate profile of the Aeromonas hydrophila CphA metallo-β-lactamase by site-directed mutagenesis. / Bebrone, Carine; Anne, Christine; De Vriendt, Kris; Devreese, Bart; Rossolini, Gian Maria; Van Beeumen, Jozef; Frère, Jean Marie; Galleni, Moreno.

In: Journal of Biological Chemistry, Vol. 280, No. 31, 05.08.2005, p. 28195-28202.

Research output: Contribution to journalArticle

Bebrone, C, Anne, C, De Vriendt, K, Devreese, B, Rossolini, GM, Van Beeumen, J, Frère, JM & Galleni, M 2005, 'Dramatic broadening of the substrate profile of the Aeromonas hydrophila CphA metallo-β-lactamase by site-directed mutagenesis', Journal of Biological Chemistry, vol. 280, no. 31, pp. 28195-28202. https://doi.org/10.1074/jbc.M414052200
Bebrone, Carine ; Anne, Christine ; De Vriendt, Kris ; Devreese, Bart ; Rossolini, Gian Maria ; Van Beeumen, Jozef ; Frère, Jean Marie ; Galleni, Moreno. / Dramatic broadening of the substrate profile of the Aeromonas hydrophila CphA metallo-β-lactamase by site-directed mutagenesis. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 31. pp. 28195-28202.
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