Identification of residues critical for the human coronavirus 229E receptor function of human aminopeptidase N

J Gen Virol. 1997 Nov;78 ( Pt 11):2795-802. doi: 10.1099/0022-1317-78-11-2795.

Abstract

Aminopeptidase N (APN) is the major cell surface receptor for group 1 coronaviruses. In this study, we have isolated and characterized a feline APN cDNA and shown that the transfection of human embryonic kidney cells with this cDNA renders them susceptible to infection with the feline coronavirus feline infectious peritonitis virus, the human coronavirus (HCV) 229E and the porcine coronavirus porcine transmissible gastroenteritis virus. By using chimeric APN genes, assembled from porcine and feline sequences, we have shown that, analogously to the human APN protein, a region within the amino-terminal part of the feline APN protein (encompassing amino acids 132-295) is essential for its HCV 229E receptor function. Furthermore, by comparing the relevant feline, human and porcine APN sequences, we were able to identify a hypervariable stretch of eight amino acids that are more closely related in the feline and human APN proteins than in the porcine APN molecule. Using PCR-directed mutagenesis, we converted this stretch of amino acids within the porcine APN molecule to the corresponding residues of the human APN molecule. These changes were sufficient to convert porcine APN into a functional receptor for HCV 229E and thus define a small number of residues that are critically important for the HCV 229E receptor function of human APN.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites / genetics
  • CD13 Antigens / genetics*
  • CD13 Antigens / metabolism
  • Cats
  • Cell Line
  • Cloning, Molecular
  • Coronavirus / physiology*
  • Coronavirus 229E, Human*
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Polymerase Chain Reaction
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Virus Replication / genetics*

Substances

  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • CD13 Antigens