Publication: Identification of important n-linked glycosylation sites in the hemagglutinin protein and their functional impact on DC-SIGN mediated avian influenza H5N1 infection
Issued Date
2021-01-02
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ISSN
14220067
16616596
16616596
Other identifier(s)
2-s2.0-85099402693
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Mahidol University
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SCOPUS
Bibliographic Citation
International Journal of Molecular Sciences. Vol.22, No.2 (2021), 1-22
Suggested Citation
Zih Syuan Yang, Szu Wei Huang, Wen Hung Wang, Chih Yen Lin, Chu Feng Wang, Aspiro Nayim Urbina, Arunee Thitithanyanont, Sung Pin Tseng, Po Liang Lu, Yen Hsu Chen, Sheng Fan Wang Identification of important n-linked glycosylation sites in the hemagglutinin protein and their functional impact on DC-SIGN mediated avian influenza H5N1 infection. International Journal of Molecular Sciences. Vol.22, No.2 (2021), 1-22. doi:10.3390/ijms22020743 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/76310
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Title
Identification of important n-linked glycosylation sites in the hemagglutinin protein and their functional impact on DC-SIGN mediated avian influenza H5N1 infection
Abstract
DC-SIGN, a C-type lectin mainly expressed in dendritic cells (DCs), has been reported to mediate several viral infections. We previously reported that DC-SIGN mediated H5N1 influenza A virus (AIVs) infection, however, the important DC-SIGN interaction with N-glycosylation sites remain unknown. This study aims to identify the optimal DC-SIGN interacting N-glycosylation sites in HA proteins of H5N1-AIVs. Results from NetNGlyc program analyzed the H5 hemagglutinin sequences of isolates during 2004–2020, revealing that seven and two conserved N-glycosylation sites were detected in HA1 and HA2 domain, respectively. A lentivirus pseudotyped A/Vietnam/1203/04 H5N1 envelope (H5N1-PVs) was generated which displayed an abundance of HA5 proteins on the virions via immuno-electron microscope observation. Further, H5N1-PVs or reverse-genetics (H5N1-RG) strains carrying a serial N-glycosylated mutation was generated by site-directed mutagenesis assay. Human recombinant DC-SIGN (rDC-SIGN) coated ELISA showed that H5N1-PVs bound to DC-SIGN, however, mutation on the N27Q, N39Q, and N181Q significantly reduced this binding (p < 0.05). Infectivity and capture assay demonstrated that N27Q and N39Q mutations significantly ameliorated DC-SIGN mediated H5N1 infection. Furthermore, combined mutations (N27Q&N39Q) significantly waned the interaction on either H5N1-PVs or-RG infection in cis and in trans (p < 0.01). This study concludes that N27 and N39 are two essential N-glycosylation contributing to DC-SIGN mediating H5N1 infection.