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FEMS microbiology letters2003; 222(1); 69-74; doi: 10.1016/S0378-1097(03)00222-2

CNE, a collagen-binding protein of Streptococcus equi.

Abstract: Streptococcus equi subspecies equi is an important horse pathogenic bacterium causing a serious disease called strangles. Using bioinformatics we identified a gene denoted cne (gene encoding collagen-binding protein from S. equi) coding for a novel potential virulence factor of this species called protein CNE. The protein is composed of 657 amino acids and has the typical features found in cell surface-anchored proteins in Gram-positive bacteria. CNE displays amino acid sequence similarities to the previously well-studied collagen-binding protein CNA from Staphylococcus aureus, a proven virulence factor in septic arthritis. Based on similarity to CNA the structure of the mature CNE protein can be divided into an N-terminal A domain and a C-terminal B domain. The highest similarity between CNA and CNE is found in the A domains. The A domain in CNA is known to be the collagen-binding domain. Two parts of cne were amplified using polymerase chain reaction (PCR) and ligated into an expression vector, and recombinant CNE proteins were produced in Escherichia coli. The purified CNE proteins were shown to display collagen-binding activity in a Western ligand blot and to inhibit collagen binding to cells of subsp. equi and to CNE-coated microtitre wells. Furthermore, the A domain of CNE was sufficient for binding collagen, and was shown to compete for the same site on collagen as CNA in inhibition studies. Using PCR, the cne gene was detected in all studied strains of subsp. equi and S. equi subsp. zooepidemicus.
Publication Date: 2003-05-22 PubMed ID: 12757948DOI: 10.1016/S0378-1097(03)00222-2Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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The study investigates a potential virulence factor in the bacterium Streptococcus equi, which causes a serious horse disease called strangles. Through bioinformatics, the researchers identified a gene that codes for a protein called CNE, which binds to collagen. Testing showed that this protein was capable of inhibiting collagen binding in multiple contexts.

About the Study

  • The research focuses on the bacterium Streptococcus equi, which is responsible for a severe equine disease known as strangles.
  • Using bioinformatics techniques, the researchers isolated a particular gene known as cne — this codes for a collagen-binding protein called CNE, that the team hypothesize may play a role in the bacterium’s virulence.

The CNE Protein

  • This protein, made up of 657 amino acids, bears similar features to other proteins found on the surface of Gram-positive bacteria.
  • CNE shows similarities to another collagen-binding protein, CNA, found in Staphylococcus aureus — another pathogenic bacterium known to be a key player in septic arthritis.
  • The researchers were able to distinguish two primary domains in CNE’s structure, similar to CNA: the N-terminal (or A domain), and the C-terminal (or B domain). The highest similarities between the two aforementioned proteins were found in their A domains, known to be responsible for collagen binding.

Experimentation and Findings

  • Parts of the cne gene were amplified via polymerase chain reaction (PCR) and subsequently inserted into an “expression vector”. This allowed the researchers to produce copies of the CNE protein in Escherichia coli.
  • These recopied CNE proteins, once purified, exhibited the ability to bind to collagen in a Western ligand blot.
  • The experiments showed that the CNE protein prevented (or inhibited) collagen binding to cells of the Streptococcus equi bacterium.
  • Further exploration of the A domain of the CNE protein demonstrated that it was sufficient for collagen binding and that it competed for the same collagen binding site as CNA in the inhibition studies.
  • Finally, the cne gene was detected in each strain of Streptococcus equi and Streptococcus equi subsp. zooepidemicus tested via PCR.

Cite This Article

APA
Lannergård J, Frykberg L, Guss B. (2003). CNE, a collagen-binding protein of Streptococcus equi. FEMS Microbiol Lett, 222(1), 69-74. https://doi.org/10.1016/S0378-1097(03)00222-2

Publication

ISSN: 0378-1097
NlmUniqueID: 7705721
Country: England
Language: English
Volume: 222
Issue: 1
Pages: 69-74

Researcher Affiliations

Lannergård, Jonas
  • Department of Microbiology, Swedish University of Agricultural Sciences, PO Box 7025, 750 07 Uppsala, Sweden.
Frykberg, Lars
    Guss, Bengt

      MeSH Terms

      • Animals
      • Bacterial Proteins / genetics
      • Bacterial Proteins / metabolism
      • Collagen / metabolism
      • Horse Diseases / microbiology
      • Horses
      • Staphylococcus aureus
      • Streptococcal Infections / microbiology
      • Streptococcal Infections / veterinary
      • Streptococcus equi / genetics
      • Streptococcus equi / metabolism
      • Streptococcus equi / pathogenicity
      • Virulence

      Citations

      This article has been cited 27 times.
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