Binding characterization of determinants in porcine aminopeptidase N, the cellular receptor for transmissible gastroenteritis virus.
Four truncated porcine aminopeptidase N (pAPN, a cellular receptor for porcine coronaviruses) proteins were expressed in prokaryotic cells. The recognizing of a specific serum against pAPN to these proteins was investigated by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The binding ability of the proteins to transmissible gastroenteritis virus (TGEV), a porcine coronavirus, was analyzed by ELISA. The inhibitory effect of these proteins to cell infection by TGEV was analyzed using plaque assays. Our data indicate that three truncated pAPNs positively reacted with the specific antiserum and the major binding regions of pAPN were limited in regions 36aa-223aa, 349aa-591aa and 592-963aa. The proteins showed discrepant binding activity to either pAPN antibody or TGE virions. Moreover, the truncated proteins blocked the infection of cells by TGEV to different extent. The results suggest that the major antibody-binding domains of pAPN may associate with the receptor-binding determinants. The role of APN is discussed in the context of virus receptor usage.
Ren
Xiaofeng
X
College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, 150030 Harbin, China. rxfemail@yahoo.com.cn
Li
Guangxing
G
Liu
Boqi
B
D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics; D001665:Binding Sites / Q000235:genetics*; D018826:CD13 Antigens / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D015151:Immunoblotting; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics; D013552:Swine / Q000235:genetics* / Q000378:metabolism; D005760:Transmissible gastroenteritis virus / Q000378:metabolism*; D010948:Viral Plaque Assay; D014771:Virion / Q000235:genetics
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D018826:CD13 Antigens