Structural insight into the bacterial mucinase StcE essential to adhesion and immune evasion during enterohemorrhagic E. coli infection

Structure. 2012 Apr 4;20(4):707-17. doi: 10.1016/j.str.2012.02.015. Epub 2012 Apr 3.

Abstract

Mucin glycoproteins with large numbers of O-linked glycosylations comprise the mucosal barrier lining the mammalian gastrointestinal tract from mouth to gut. A critical biological function of mucins is to protect the underlying epithelium from infection. Enterohemorrhagic Escherichia coli (EHEC), the mediator of severe food- and water-borne disease, can breach this barrier and adhere to intestinal cells. StcE, a ∼100 kDa metalloprotease secreted by EHEC, plays a pivotal role in remodeling the mucosal lining during infection. To obtain mechanistic insight into its function, we have determined the structure of StcE. Our data reveal a dynamic, multidomain architecture featuring an unusually large substrate-binding cleft and a prominent polarized surface charge distribution highly suggestive of an electrostatic role in substrate targeting. The observation of key conserved motifs in the active site allows us to propose the structural basis for the specific recognition of α-O-glycan-containing substrates. Complementary biochemical analysis provides further insight into its distinct substrate specificity and binding stoichiometry.

Publication types

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

MeSH terms

  • Bacterial Adhesion / genetics*
  • Bacterial Adhesion / immunology
  • Carbohydrate Conformation
  • Catalytic Domain
  • Crystallography, X-Ray
  • Enterohemorrhagic Escherichia coli / chemistry*
  • Enterohemorrhagic Escherichia coli / immunology
  • Enterohemorrhagic Escherichia coli / metabolism
  • Escherichia coli Infections / immunology
  • Escherichia coli Infections / microbiology
  • Escherichia coli Proteins / chemistry*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Humans
  • Immune Evasion
  • Intestinal Mucosa / immunology
  • Intestinal Mucosa / microbiology
  • Metalloendopeptidases / chemistry*
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism
  • Models, Molecular
  • Mucins / chemistry*
  • Mucins / metabolism
  • Protein Binding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Static Electricity
  • Stereoisomerism
  • Substrate Specificity

Substances

  • Escherichia coli Proteins
  • Mucins
  • Recombinant Proteins
  • Metalloendopeptidases
  • StcE protein, E coli

Associated data

  • PDB/3UJZ
  • PDB/4DNY