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Identification of Porcine haemagglutinating encephalomyelitis virus receptor in PK cell membranes

Published online by Cambridge University Press:  27 June 2008

Lu Hui-Jun
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
He Wen-Qi
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Song De-Guang
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Liu Li-Guo
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Chang Ling-Zhu
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Li Zhi-Ping
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Chen Ke-Yan
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Gao Feng*
Affiliation:
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
*
*Corresponding author. E-mail: [email protected]

Abstract

To identify Porcine haemagglutinating encephalomyelitis virus (HEV) 67N receptor in porcine kidney (PK) cell membranes, the S1 protein of HEV was expressed in Pichia pastoris and purified by Ni2+ affinity chromatograph. Polyclonal antibodies to HEV were prepared by immunizing rabbits by injecting the purified S1 protein four times. After SDS–polyacrylamide gel electrophoresis (SDS–PAGE), the PK cell membrane proteins were transferred on to nitrocellulose membrane. A virus overlay protein binding assay (VOPBA) was performed using the recombinant S1 protein to identify the protein binding receptor, HEV-S1. The result showed that HEV-S1 protein bound to one band (about 90 kDa) in PK cell membranes. This result is very important for the study of the pathogenic mechanism of HEV.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(1): 20–23

References

Callagher, TM and Buchmeier, MJ (2001) Coronavirus spike proteins in viral entry and pathogenesis. Viiroloy 279(2): 371374.CrossRefGoogle Scholar
Chang, CN, Hsu, FS, Shen, YM and Yen, CC (1978) A serological survey on hemagglutinating encephalomyelitis virus infection in pigs of Taiwan Sugar Corporation. Annual Research Report of Animal Research Institute. Taiwan: Taiwan Sugar Corporation, pp. 157163.Google Scholar
Gao, F, Ding, Z, Zhang, ML, et al. (2000) Outbreak of piglet coronavirus encephalomyelitis in three farms. Chinese Journal of Preventive Veterinary Medicine 22(2): 151152.Google Scholar
Guo, AZ and Lu, CP (2000) Identification of canine distemper virus receptor in cell lines. Chinese Journal of Virology 16(2): 155157.Google Scholar
He, WQ, Lu, HJ, Geng, BC, et al. (2005) Serological investigation of antibody against hemagglutinating encephalomyelitis virus in pigs. Chinese Journal of Veterinary Science and Technology 35(9): 739741.Google Scholar
Hirano, N, Suzuki, Y and Haga, S (1999) Pigs with highly prevalent antibodies to human coronavirus and swine haemagglutinating encephalomyelitis virus in the Tohoku District of Japan. Epidemiology and Infection 122: 545551.CrossRefGoogle ScholarPubMed
Krueger, DK, Kelly, SM, Lewicki, DN, Ruffolo, R and Gallagher, TM (2001) Variations in disparate regions of the murine coronavirus spike protein impact the initiation of membrane fusion. Journal of Virology 75(6): 27922802.CrossRefGoogle ScholarPubMed
Matsnyama, S and Taguchi, F (2002) Communication between S1N330 and a region in S2 of murine coronavirus spike protein is important for virus entry into cells expressing CEACAM1b receptor. Virology 295(1): 160171.CrossRefGoogle Scholar
McFerren, JB, Clarke, JK, Cornner, TJ and Knox, ER (1971) Serological evidence of the presence of hemaglutinating encephalomyelitis virus in Northern Ireland. Veterinary Record 88: 339340.CrossRefGoogle Scholar
Quiroga, MA, Cappuccio, J, Piñeyro, P, et al. (2008) Hemagglutinating encephalomyelitis coronavirus infection in pigs, Argentina. Emerging Infectious Diseases 14(3): 484486.CrossRefGoogle ScholarPubMed
Roe, CK and Alexander, TJL (1958) A disease of nursing pigs previously unreported in Ontario. Canadian Journal of Veterinary Medicine 22: 305307.Google ScholarPubMed
Saif, LJ (1993) Coronavirus immunogens. Veterinary Microbiology 37(3–4): 285297.CrossRefGoogle ScholarPubMed
Sasseville, AM, Gelinas, AM, Sawyer, N, Boutin, M and Dea, S (2001) Biological and molecular characteristics of an HEV isolate associated with recent acute outbreaks of encephalomyelitis in Quebec pig farms. Advances in Experimental Medicine and Biology 494: 5762.CrossRefGoogle ScholarPubMed
Taguchi, F and Shimazaki, YK (2000) Functional analysis of an epitope in the S2 subunit of the murine coronavirus spike protein: invovlement in fusion activity. Journal of General Virology 81(12): 28672871.CrossRefGoogle Scholar
Weiss, SR and Navas-Martin, S (2005) Coronavirus pathogenesis and the emerging pathogen severe acute respiratory syndrome coronavirus. Microbiology and Molecular Biology Review 69(4): 635664.CrossRefGoogle ScholarPubMed
Wentworth, DE and Holmes, KV (2001) Molecular determinants of species specificity in the coronavirus receptor aminopeptidase N (CD13): influence of N-linked glycosylation. Journal of Virology 75: 97419752.CrossRefGoogle ScholarPubMed
Xu, YX, Zhou, XF and Liu, LD (2000) Molecular Virology. Hubei: Science & Technology Press, pp. 324330.Google Scholar
Yin, Z and Liu, J H (1997) Animal Virology. Peking: Science Press, pp. 690692.Google Scholar