Related Experiment Video
Updated: Jan 31, 2026

A Miniaturized Glycan Microarray Assay for Assessing Avidity and Specificity of Influenza A Virus Hemagglutinins
Published on: May 29, 2016
Fluorescent Trimeric Hemagglutinins Reveal Multivalent Receptor Binding Properties
Nikoloz Nemanichvili1, Ilhan Tomris2, Hannah L Turner3
1Pathology Division, Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, 3584, CL, Utrecht, the Netherlands.
Influenza hemagglutinin (HA) proteins fused with sfGFP visualize host cell interactions. Avian-specific HA requires multimerization for binding on arrays, unlike human-specific HA, suggesting distinct binding mechanisms.
Area of Science:
- Virology
- Biochemistry
- Molecular Biology
Background:
- Influenza A virus hemagglutinin (HA) mediates host cell entry via multivalent binding to sialylated glycans.
- Current receptor binding assays often use viruses or precomplexed HA trimers, limiting direct visualization of HA-glycan interactions.
Purpose of the Study:
- To develop a method for direct visualization of influenza HA-receptor interactions.
- To investigate the multivalent binding properties of human- and avian-specific HA using a novel fusion protein.
Main Methods:
- Fusion of superfolder green fluorescent protein (sfGFP) to the C-terminus of trimeric HA.
- Utilizing glycan arrays and tissue staining to analyze HA-sfGFP binding.
- Comparing binding of human- and avian-specific HA-sfGFP with and without antibody-mediated multimerization.
Main Results:
- Human-specific HA-sfGFP bound to glycan arrays as a free trimer.
- Avian-specific HA-sfGFP required antibody-mediated multimerization for glycan array binding.
- Multimerization was not necessary for avian-specific HA-sfGFP binding to tissues.
- Differences in binding may be due to glycan structure and presentation on arrays versus cell surfaces.
Conclusions:
- The sfGFP fusion allows direct visualization of HA-glycan interactions.
- Human- and avian-specific HA exhibit distinct binding requirements influenced by glycan presentation.
- Cell surface presentation of glycans facilitates avian-specific HA binding, overcoming geometrical constraints observed on arrays.
More Related Videos
10:13Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
Published on: June 9, 2017
14:09Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell
Published on: August 4, 2015
Related Concept Videos
Internal Receptors
Ligand Binding and Linkage
The Equilibrium Binding Constant and Binding Strength
The Equilibrium Binding Constant and Binding Strength
Receptor-mediated Endocytosis
G-protein Coupled Receptors