Crystallization and preliminary X-ray diffraction analysis of CfaE, the adhesive subunit of the CFA/I fimbriae from human enterotoxigenic Escherichia coli

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Feb 1;62(Pt 2):121-4. doi: 10.1107/S1744309105043198. Epub 2006 Jan 27.

Abstract

Enterotoxigenic Escherichia coli (ETEC) represents a formidable food and waterborne diarrheal disease threat of global importance. The first step in ETEC pathogenesis is bacterial attachment to small-intestine epithelial cells via adhesive fimbriae, many of which are genetically related to the prototype colonization factor antigen I (CFA/I). The minor fimbrial subunit CfaE is required for initiation of CFA/I fimbrial assembly and mediates bacterial attachment to host cell-surface receptors. A donor-strand complemented variant of CfaE (dscCfaE) was expressed with a hexahistidine tag, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion method. X-ray diffraction data sets were collected to 2.4 A resolution for both native and derivatized crystals and showed the symmetry of space group P6(2)22, with unit-cell parameters a = b = 142.9, c = 231.9 A. Initial phases were derived from the SIRAS approach and electron density showed two molecules in the crystallographic asymmetric unit. Sequence assignments were aided by anomalous signals from the selenium of an SeMet-derivatized crystal and from S atoms of a native crystal.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Adhesion / physiology
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / pathogenicity*
  • Escherichia coli / physiology
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / physiology
  • Fimbriae Proteins / chemistry*
  • Fimbriae Proteins / physiology
  • Fimbriae, Bacterial / chemistry*
  • Fimbriae, Bacterial / physiology
  • Humans
  • Protein Subunits / chemistry*
  • Protein Subunits / physiology

Substances

  • Escherichia coli Proteins
  • Protein Subunits
  • colonization factor antigens
  • Fimbriae Proteins