Crystallization and preliminary diffraction analysis of Wzi, a member of the capsule export and assembly pathway in Escherichia coli

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Dec 1;66(Pt 12):1621-5. doi: 10.1107/S1744309110040546. Epub 2010 Nov 26.

Abstract

External polysaccharide capsules provide a physical barrier that is employed by many species of bacteria for the purposes of host evasion and persistence. Wzi is a 53 kDa outer membrane β-barrel protein that is thought to play a role in the attachment of group 1 capsular polysaccharides to the cell surface. The purification and crystallization of an Escherichia coli homologue of Wzi is reported and diffraction data from native and selenomethionine-incorporated protein crystals are presented. Crystals of C-terminally His6-tagged Wzi diffracted to 2.8 Å resolution. Data processing showed that the crystals belonged to the orthorhombic space group C222, with unit-cell parameters a=128.8, b=152.8, c=94.4 Å, α=β=γ=90°. A His-tagged selenomethionine-containing variant of Wzi has also been crystallized in the same space group and diffraction data have been recorded to 3.8 Å resolution. Data processing shows that the variant crystal has similar unit-cell parameters to the native crystal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Capsules / metabolism*
  • Bacterial Outer Membrane Proteins / chemistry*
  • Biosynthetic Pathways*
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / chemistry*
  • Escherichia coli Proteins / chemistry*
  • Selenomethionine / chemistry
  • X-Ray Diffraction*

Substances

  • Bacterial Outer Membrane Proteins
  • Escherichia coli Proteins
  • Wzi protein, E coli
  • Selenomethionine