Next Article in Journal
The Interplay between Angiopoietin-Like Proteins and Adipose Tissue: Another Piece of the Relationship between Adiposopathy and Cardiometabolic Diseases?
Previous Article in Journal
Arsenite Inhibits Tissue-Type Plasminogen Activator Synthesis through NRF2 Activation in Cultured Human Vascular Endothelial EA.hy926 Cells
Previous Article in Special Issue
Extended Interactions between HIV-1 Viral RNA and tRNALys3 Are Important to Maintain Viral RNA Integrity
Open AccessArticle

Identification of Important N-Linked Glycosylation Sites in the Hemagglutinin Protein and Their Functional Impact on DC-SIGN Mediated Avian Influenza H5N1 Infection

1
Center for Tropical Medicine and Infectious Disease, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
2
Department of Medical Laboratory Science and Biotechnology, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
3
Model Development Section, Basic Research Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA
4
Division of Infectious Disease, Department of Internal Medicine, Kaohsiung Medical, University Hospital, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
5
Clinical Microbiology Laboratory, Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
6
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
7
Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
*
Author to whom correspondence should be addressed.
These authors contributed equally to this study.
Int. J. Mol. Sci. 2021, 22(2), 743; https://doi.org/10.3390/ijms22020743
Received: 15 December 2020 / Revised: 5 January 2021 / Accepted: 11 January 2021 / Published: 13 January 2021
(This article belongs to the Special Issue Virus Replication)
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. View Full-Text
Keywords: DC-SIGN; H5N1; N27Q; N39Q; hemagglutinin; N-linked glycosylation; infection DC-SIGN; H5N1; N27Q; N39Q; hemagglutinin; N-linked glycosylation; infection
Show Figures

Figure 1

MDPI and ACS Style

Yang, Z.-S.; Huang, S.-W.; Wang, W.-H.; Lin, C.-Y.; Wang, C.-F.; Urbina, A.N.; Thitithanyanont, A.; Tseng, S.-P.; Lu, P.-L.; Chen, Y.-H.; Wang, S.-F. Identification of Important N-Linked Glycosylation Sites in the Hemagglutinin Protein and Their Functional Impact on DC-SIGN Mediated Avian Influenza H5N1 Infection. Int. J. Mol. Sci. 2021, 22, 743. https://doi.org/10.3390/ijms22020743

AMA Style

Yang Z-S, Huang S-W, Wang W-H, Lin C-Y, Wang C-F, Urbina AN, Thitithanyanont A, Tseng S-P, Lu P-L, Chen Y-H, Wang S-F. 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. 2021; 22(2):743. https://doi.org/10.3390/ijms22020743

Chicago/Turabian Style

Yang, Zih-Syuan; Huang, Szu-Wei; Wang, Wen-Hung; Lin, Chih-Yen; Wang, Chu-Feng; Urbina, Aspiro N.; Thitithanyanont, Arunee; Tseng, Sung-Pin; Lu, Po-Liang; Chen, Yen-Hsu; Wang, Sheng-Fan. 2021. "Identification of Important N-Linked Glycosylation Sites in the Hemagglutinin Protein and Their Functional Impact on DC-SIGN Mediated Avian Influenza H5N1 Infection" Int. J. Mol. Sci. 22, no. 2: 743. https://doi.org/10.3390/ijms22020743

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop