The Class D beta-lactamase family: residues governing the maintenance and diversity of function.

摘要

Class D beta-lactamases, a major source of bacterial resistance to beta-lactam antibiotic therapies, represent a distinct subset of the beta-lactamase superfamily. They share a serine hydrolase mechanism with Classes A/C vs. Class B. Further understanding of their sequence-structure-function relationships would benefit efforts to design a new generation of antibiotics as well as to predict evolutionary mechanisms in response to such therapies. Here we describe analyses based on our high-resolution multiple sequence alignment and phylogenetic tree of approximately 80 Class D beta-lactamases that leverage several 3D structures of these enzymes. We observe several sequence clusters on the phylogenetic tree, some that are species specific while others include several species from alpha-, beta- and gamma-proteobacteria. Residues characteristic of a specific cluster were identified and shown to be located just outside the active site, possibly modulating the function of the catalytic residues to facilitate reactions with specific types of beta-lactams. Most significant was the discovery of a likely disulfide bond in a large group composed of alpha-, beta- and gamma-proteobacteria that would contribute to enzyme stability and hence bacterial viability under antibiotic assault. A network of co-evolving residues was identified which suggested the importance of maintaining a surface for binding a highly conserved Phe69.