Kinetic Studies on CphA Mutants Reveal the Role of the P158-P172 Loop in Activity versus Carbapenems

Antimicrob Agents Chemother. 2016 Apr 22;60(5):3123-6. doi: 10.1128/AAC.01703-15. Print 2016 May.

Abstract

Site-directed mutagenesis of CphA indicated that prolines in the P158-P172 loop are essential for the stability and the catalytic activity of subclass B2 metallo-β-lactamases against carbapenems. The sequential substitution of proline led to a decrease of the catalytic efficiency of the variant compared to the wild-type (WT) enzyme but also to a higher affinity for the binding of the second zinc ion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Binding Sites
  • Carbapenems / pharmacology*
  • Kinetics
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Proline / chemistry
  • Proline / metabolism
  • Sequence Analysis, Protein
  • Structure-Activity Relationship
  • Substrate Specificity / genetics
  • Substrate Specificity / physiology
  • Zinc / pharmacology
  • beta-Lactamases / chemistry
  • beta-Lactamases / genetics
  • beta-Lactamases / metabolism*

Substances

  • Bacterial Proteins
  • Carbapenems
  • Proline
  • beta-Lactamases
  • Zinc