A comparative study of class-D beta-lactamases

Biochem J. 1993 Jun 1;292 ( Pt 2)(Pt 2):555-62. doi: 10.1042/bj2920555.

Abstract

Three class-D beta-lactamases (OXA2, OXA1 and PSE2) were produced and purified to protein homogeneity. 6 beta-Iodopenicillanate inactivated the OXA2 enzyme without detectable turnover. Labelling of the same beta-lactamase with 6 beta-iodo[3H]penicillanate allowed the identification of Ser-70 as the active-site serine residue. In agreement with previous reports, the apparent M(r) of the OXA2 enzyme as determined by molecular-sieve filtration, was significantly higher than that deduced from the gene sequence, but this was not due to an equilibrium between a monomer and a dimer. The heterogeneity of the OXA2 beta-lactamase on ion-exchange chromatography contrasted with the similarity of the catalytic properties of the various forms. A first overview of the enzymic properties of the three 'oxacillinases' is presented. With the OXA2 enzyme, 'burst' kinetics, implying branched pathways, seemed to prevail with many substrates.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Catalysis
  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Cloning, Molecular
  • Kinetics
  • Penicillanic Acid / metabolism
  • Plasmids
  • Serine / metabolism
  • Substrate Specificity
  • beta-Lactamase Inhibitors
  • beta-Lactamases / chemistry
  • beta-Lactamases / isolation & purification
  • beta-Lactamases / metabolism*

Substances

  • beta-Lactamase Inhibitors
  • Serine
  • Penicillanic Acid
  • 6-iodopenicillanic acid
  • beta-Lactamases