Cloning, expression, purification and preliminary X-ray diffraction studies of a novel AB₅ toxin

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Aug;69(Pt 8):912-5. doi: 10.1107/S1744309113018927. Epub 2013 Jul 27.

Abstract

AB₅ toxins are key virulence factors found in a range of pathogenic bacteria. AB₅ toxins consist of two components: a pentameric B subunit that targets eukaryotic cells by binding to glycans located on the cell surface and a catalytic A subunit that disrupts host cellular function following internalization. To date, the A subunits of AB₅ toxins either have RNA-N-glycosidase, ADP-ribosyltransferase or serine protease activity. However, it has been suggested that a novel AB₅ toxin produced by clinical isolates of Escherichia coli and Citrobacter freundii has an A subunit with metalloproteinase activity. Here, the expression, purification and crystallization of this novel AB₅ toxin from E. coli (EcxAB) and the collection of X-ray data to 1.9 Å resolution are reported.

Keywords: AB5 toxins; Escherichia coli; co-expression; proteases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Toxins / biosynthesis
  • Bacterial Toxins / chemistry
  • Bacterial Toxins / genetics*
  • Cloning, Molecular* / methods
  • Escherichia coli Proteins / biosynthesis
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics*
  • Gene Expression Regulation, Bacterial*
  • Protein Subunits / chemistry
  • Protein Subunits / genetics
  • X-Ray Diffraction

Substances

  • Bacterial Toxins
  • Escherichia coli Proteins
  • Protein Subunits