Three-dimensional structure of Wza, the protein required for translocation of group 1 capsular polysaccharide across the outer membrane of Escherichia coli

J Biol Chem. 2004 Jul 2;279(27):28227-32. doi: 10.1074/jbc.M402913200. Epub 2004 Apr 16.

Abstract

Wza is a highly conserved multimeric outer membrane protein complex required for the surface expression of the serotype K30 group 1 capsular polysaccharide in Escherichia coli. Here we present the first three-dimensional structure of this type of polysaccharide exporter at a 15.5-A resolution obtained using single particle averaging on a dataset of cryo-negatively stained protein. Previous structural studies on purified Wza have revealed a homo-oligomeric ring structure that is most probably composed of eight subunits. Symmetry analysis of the three-dimensional structure combined with biochemical two- and three-dimensional crystallographic data strongly suggest that Wza is an octameric complex with a C4 quasi-rotational symmetry and is organized as a tetramer of dimeric subunits. Wza is best described as a stack of two 4-A high rings with differing diameters providing a mushroom-like aspect from the side. The larger ring has a distinctive square shape with a diameter of 115 A, whereas the smaller is almost circular with a diameter of 90 A. In the center of the complex and enclosed by the four symmetrical arms is a small elliptical cagelike cavity of approximately 40 A in diameter. The central cavity is effectively sealed at the top and bottom of the complex but has small inter-arm holes when viewed from the side. We discuss the structure of this complex and implications in the surface translocation of cell-surface polysaccharide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Outer Membrane Proteins / chemistry*
  • Biological Transport
  • Cryoelectron Microscopy
  • Crystallization
  • Crystallography, X-Ray
  • Dimerization
  • Escherichia coli Proteins / chemistry*
  • Image Processing, Computer-Assisted
  • Microscopy, Electron
  • Models, Molecular
  • Polysaccharides / chemistry*
  • Protein Conformation
  • Protein Structure, Tertiary
  • Protein Transport

Substances

  • Bacterial Outer Membrane Proteins
  • Escherichia coli Proteins
  • Polysaccharides
  • Wza protein, E coli