Protocol

SARS- and Other Coronaviruses

Volume 454 of the series Methods in Molecular Biology pp 319-329

Date:

Identification of Sugar Residues Involved in the Binding of TGEV to Porcine Brush Border Membranes

  • Christel Schwegmann-WesselsAffiliated withInstitute for Virology, University of Veterinary Medicine Hannover
  • , Georg HerrlerAffiliated withInstitute for Virology, University of Veterinary Medicine Hannover

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Abstract

Coronaviruses most often infect the respiratory or intestinal tract. Transmissible gastroenteritis virus (TGEV), a group 1 coronavirus, infects the porcine small intestine. Piglets up to the age of 3 weeks die from diarrhea caused by the viral gastroenteritis unless they are protected by antibodies. In addition to the cellular receptor, porcine aminopeptidase N, the TGEV spike protein binds to sialic acid residues. We have shown that the sialic acid binding activity mediates the binding of TGEV to a mucin-like glycoprotein present in porcine brush border membranes. This was shown by performing a virus overlay binding assay with proteins obtained from brush border membranes by lectin precipitation. Because of the reactivity with specific lectins we assume that the recognized glycoprotein has the characteristics of a mucin.

Key Words

transmissible gastroenteritis virus sialic acid binding activity brush border membranes piglets lectin precipitation virus overlay binding assay