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Sd a antigen expression reduces paramyxovirus HN and influenza HA binding to glycoprotein receptors. (A) <t>LAMP1</t> and LAMP1-Sd a glycoproteins were loaded onto the BLI sensor at comparable levels. (B) Lectin characterization of LAMP1 and LAMP1-Sd a using DBA and MAL I. MAL I recognizes Neu5Acα2-3Galβ1-4GlcNAc (a trisaccharide common in N-glycans). (C) BLI analysis to determine the binding kinetics of nanoparticles displaying paramyxovirus HN or influenza HA glycoproteins to LAMP1 and LAMP1-Sd a . Binding was assessed using an established HN-Ni NTA nanoparticle (HN-NPs) system using 3.5×10 10 HN-NPs or HA-NPs per well. Binding of NDV HN-NPs was additionally tested in the presence of site-I specific inhibitor BCX2798 (BCX). Each experiment was conducted at least twice, with representative results displayed here.
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Sd a antigen expression reduces paramyxovirus HN and influenza HA binding to glycoprotein receptors. (A) <t>LAMP1</t> and LAMP1-Sd a glycoproteins were loaded onto the BLI sensor at comparable levels. (B) Lectin characterization of LAMP1 and LAMP1-Sd a using DBA and MAL I. MAL I recognizes Neu5Acα2-3Galβ1-4GlcNAc (a trisaccharide common in N-glycans). (C) BLI analysis to determine the binding kinetics of nanoparticles displaying paramyxovirus HN or influenza HA glycoproteins to LAMP1 and LAMP1-Sd a . Binding was assessed using an established HN-Ni NTA nanoparticle (HN-NPs) system using 3.5×10 10 HN-NPs or HA-NPs per well. Binding of NDV HN-NPs was additionally tested in the presence of site-I specific inhibitor BCX2798 (BCX). Each experiment was conducted at least twice, with representative results displayed here.
Human Ace2 Ectodomain With A Human Igg1 Fc Tag (Ace2 Fc), supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Sd a antigen expression reduces paramyxovirus HN and influenza HA binding to glycoprotein receptors. (A) LAMP1 and LAMP1-Sd a glycoproteins were loaded onto the BLI sensor at comparable levels. (B) Lectin characterization of LAMP1 and LAMP1-Sd a using DBA and MAL I. MAL I recognizes Neu5Acα2-3Galβ1-4GlcNAc (a trisaccharide common in N-glycans). (C) BLI analysis to determine the binding kinetics of nanoparticles displaying paramyxovirus HN or influenza HA glycoproteins to LAMP1 and LAMP1-Sd a . Binding was assessed using an established HN-Ni NTA nanoparticle (HN-NPs) system using 3.5×10 10 HN-NPs or HA-NPs per well. Binding of NDV HN-NPs was additionally tested in the presence of site-I specific inhibitor BCX2798 (BCX). Each experiment was conducted at least twice, with representative results displayed here.

Journal: Emerging Microbes & Infections

Article Title: Identification of β4GALNT2 as an anti-hPIV3 factor through genome-wide CRISPR/Cas9 library screening

doi: 10.1080/22221751.2025.2529895

Figure Lengend Snippet: Sd a antigen expression reduces paramyxovirus HN and influenza HA binding to glycoprotein receptors. (A) LAMP1 and LAMP1-Sd a glycoproteins were loaded onto the BLI sensor at comparable levels. (B) Lectin characterization of LAMP1 and LAMP1-Sd a using DBA and MAL I. MAL I recognizes Neu5Acα2-3Galβ1-4GlcNAc (a trisaccharide common in N-glycans). (C) BLI analysis to determine the binding kinetics of nanoparticles displaying paramyxovirus HN or influenza HA glycoproteins to LAMP1 and LAMP1-Sd a . Binding was assessed using an established HN-Ni NTA nanoparticle (HN-NPs) system using 3.5×10 10 HN-NPs or HA-NPs per well. Binding of NDV HN-NPs was additionally tested in the presence of site-I specific inhibitor BCX2798 (BCX). Each experiment was conducted at least twice, with representative results displayed here.

Article Snippet: Human codon-optimized cDNAs encoding the ectodomains of LAMP1 (GenScript) were cloned into the pCAGGS vector ( pCAGGS-LAMP1 ), incorporating a C-terminal Twin-Strep tag, a biotin-acceptor peptide (BAP; GGLNDIFEAQKIEWH) for site-specific biotinylation by BirA, and a 6xHis tag for purification using Ni-NTA beads [ ].

Techniques: Expressing, Binding Assay

Structural basis for Sd a -mediated disruption of hPIV3 HN-glycan interactions. (A, B) Predicted interactions between hPIV3 HN monomer and ligands using Chai Discovery ( https://lab.chaidiscovery.com ) . Panel (A) illustrates the binding of HN to 3′-sialyl-N-acetyllactosamine (3'SLN), while panel (B) depicts the interaction with the Sd a glycotope. Aromatic residues W371, F372 and W428 are shown in red. (C) Overlay of the predicted interactions, highlighting the differences between the two ligands. The interaction between GalNAc and residues C214 and R129 induces a shift in the entire glycan chain, as indicated by the red arrow. (D) Binding analysis of hPIV3 HN wild-type (WT) and W371A mutant HN-NPs to 3'S(LN) 3 was conducted using BLI, in the presence or absence of the BCX2798 inhibitor, using 3.5×10 10 HN-NPs per well. The difference in the binding curve for hPIV3 HN-NPs compared to C results from the different receptors (LAMP1 vs 3’S[LN] 3 ) being used. All experiments were performed independently in triplicate, with representative data presented.

Journal: Emerging Microbes & Infections

Article Title: Identification of β4GALNT2 as an anti-hPIV3 factor through genome-wide CRISPR/Cas9 library screening

doi: 10.1080/22221751.2025.2529895

Figure Lengend Snippet: Structural basis for Sd a -mediated disruption of hPIV3 HN-glycan interactions. (A, B) Predicted interactions between hPIV3 HN monomer and ligands using Chai Discovery ( https://lab.chaidiscovery.com ) . Panel (A) illustrates the binding of HN to 3′-sialyl-N-acetyllactosamine (3'SLN), while panel (B) depicts the interaction with the Sd a glycotope. Aromatic residues W371, F372 and W428 are shown in red. (C) Overlay of the predicted interactions, highlighting the differences between the two ligands. The interaction between GalNAc and residues C214 and R129 induces a shift in the entire glycan chain, as indicated by the red arrow. (D) Binding analysis of hPIV3 HN wild-type (WT) and W371A mutant HN-NPs to 3'S(LN) 3 was conducted using BLI, in the presence or absence of the BCX2798 inhibitor, using 3.5×10 10 HN-NPs per well. The difference in the binding curve for hPIV3 HN-NPs compared to C results from the different receptors (LAMP1 vs 3’S[LN] 3 ) being used. All experiments were performed independently in triplicate, with representative data presented.

Article Snippet: Human codon-optimized cDNAs encoding the ectodomains of LAMP1 (GenScript) were cloned into the pCAGGS vector ( pCAGGS-LAMP1 ), incorporating a C-terminal Twin-Strep tag, a biotin-acceptor peptide (BAP; GGLNDIFEAQKIEWH) for site-specific biotinylation by BirA, and a 6xHis tag for purification using Ni-NTA beads [ ].

Techniques: Disruption, Glycoproteomics, Binding Assay, Mutagenesis