The crystal structure of Escherichia coli group 4 capsule protein GfcC reveals a domain organization resembling that of Wza

Biochemistry. 2011 Jun 21;50(24):5465-76. doi: 10.1021/bi101869h. Epub 2011 May 27.

Abstract

We report the 1.9 Å resolution crystal structure of enteropathogenic Escherichia coli GfcC, a periplasmic protein encoded by the gfc operon, which is essential for assembly of group 4 polysaccharide capsule (O-antigen capsule). Presumed gene orthologs of gfcC are present in capsule-encoding regions of at least 29 genera of Gram-negative bacteria. GfcC, a member of the DUF1017 family, is comprised of tandem β-grasp (ubiquitin-like) domains (D2 and D3) and a carboxyl-terminal amphipathic helix, a domain arrangement reminiscent of that of Wza that forms an exit pore for group 1 capsule export. Unlike the membrane-spanning C-terminal helix from Wza, the GfcC C-terminal helix packs against D3. Previously unobserved in a β-grasp domain structure is a 48-residue helical hairpin insert in D2 that binds to D3, constraining its position and sequestering the carboxyl-terminal amphipathic helix. A centrally located and invariant Arg115 not only is essential for proper localization but also forms one of two mostly conserved pockets. Finally, we draw analogies between a GfcC protein fused to an outer membrane β-barrel pore in some species and fusion proteins necessary for secreting biofilm-forming exopolysaccharides.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Bacterial Capsules / chemistry
  • Bacterial Outer Membrane Proteins / chemistry*
  • Bacterial Outer Membrane Proteins / genetics
  • Base Sequence
  • Conserved Sequence
  • Crystallography, X-Ray
  • DNA, Bacterial / genetics
  • Dimerization
  • Enteropathogenic Escherichia coli / chemistry
  • Enteropathogenic Escherichia coli / genetics
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / genetics
  • Genes, Bacterial
  • Models, Molecular
  • Molecular Sequence Data
  • Operon
  • Protein Interaction Domains and Motifs
  • Protein Structure, Quaternary
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sequence Homology, Amino Acid
  • Static Electricity

Substances

  • Bacterial Outer Membrane Proteins
  • DNA, Bacterial
  • Escherichia coli Proteins
  • Recombinant Proteins
  • Wza protein, E coli