{"doi":"10.3390/ijms22020743","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.","journal":"International Journal of Molecular Sciences","year":2021,"id":177007,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":20,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9558,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":506957,"name":"Szu‐Wei Huang","orcid":"0000-0002-3813-7737","position":1,"is_corresponding":false},{"id":571427,"name":"Wen‐Hung Wang","orcid":"0000-0002-4494-1123","position":2,"is_corresponding":false},{"id":721266,"name":"C. C. Lin","orcid":null,"position":3,"is_corresponding":false},{"id":720513,"name":"Chufeng Wang","orcid":"0009-0003-4684-3564","position":4,"is_corresponding":false},{"id":721267,"name":"Aspiro Nayim Urbina","orcid":null,"position":5,"is_corresponding":false},{"id":720514,"name":"Arunee Thitithanyanont","orcid":"0000-0002-9756-4763","position":6,"is_corresponding":false},{"id":720515,"name":"Sung‐Pin Tseng","orcid":"0000-0001-8915-5268","position":7,"is_corresponding":false},{"id":510359,"name":"Po‐Liang Lu","orcid":"0000-0002-7423-6783","position":8,"is_corresponding":false},{"id":720516,"name":"Yen‐Hsu Chen","orcid":"0000-0002-8667-4844","position":9,"is_corresponding":false},{"id":506958,"name":"Sheng‐Fan Wang","orcid":"0000-0002-4183-3872","position":10,"is_corresponding":false},{"id":720512,"name":"Zih‐Syuan Yang","orcid":"0009-0007-0028-8437","position":0,"is_corresponding":true}],"reference_count":64,"raw_metadata":null,"created_at":"2026-07-18T23:47:28.018172Z","pmid":"33451024","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}