Fluorescent Trimeric Hemagglutinins Reveal Multivalent Receptor Binding Properties

Nikoloz Nemanichvili, Ilhan Tomris, Hannah L Turner, Ryan McBride, Oliver C Grant, Roosmarijn van der Woude, Mohammed H Aldosari, Roland J Pieters, Robert J Woods, James C Paulson, Geert-Jan Boons, Andrew B Ward, Monique H Verheije, Robert P de Vries

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Influenza A virus carries hundreds of trimeric hemagglutinins proteins (HA) on its viral envelope that interact with various sialylated glycans on a host cell. This interaction represents a multivalent binding event that is present in all the current receptor-binding assays, including those employing viruses or precomplexed HA trimers. To study the nature of such multivalent binding events, we fused a super folder GFP (sfGFP) to the C-terminus of trimeric HA to allow direct visualization of HA-receptor interactions without the need for additional fluorescent antibodies. The multivalent binding of the HA-sfGFP proteins was studied using glycan arrays and tissue staining. The HA-sfGFP with human-type receptor specificity was able to bind to a glycan array as the free trimer. In contrast, the HA-sfGFP with avian-type receptor specificity required multimerization by antibodies before binding to glycans on the glycan array could be observed. Interestingly, multimerization was not required for binding to tissues. The array data may be explained by the possible bivalent binding mode of a single human specific HA trimer to complex branched N-glycans, which is not possible for the avian specific HA due to geometrical constrains of the binding sites. The fact that this specificity pattern changes upon interaction with a cell surface probably represents the enhanced amount of glycan orientations and variable densities vs. those on the glycan array.

Original languageEnglish
Pages (from-to)842-856
Number of pages15
JournalJournal of Molecular Biology
Volume431
Issue number4
DOIs
Publication statusPublished - 15 Feb 2019

Keywords

  • Influenza
  • attachment protein
  • bidentate
  • multivalent
  • precomplexing

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