Glycosylation analysis of engineered H3N2 influenza A virus hemagglutinins with sequentially added historically relevant glycosylation sites

Yanming An, Jonathan Mccullers, Irina Alymova, Lisa M. Parsons, John F. Cipollo

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. The oligosaccharides of HA can contribute to HA's antigenic characteristics. After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. This glycosylation expansion has not been studied at the structural level. Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). HAs studied include HK68 and engineered forms with 1, 2, and 4 added sites. We have used: nano-LC-MSE for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. Our study reveals that 1) the majority of N-sequons are occupied at ≥ 90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. Based on this study and previous reports we provide structural insight as to how the immune system responses may differ depending on HA glycosylation.

Original languageEnglish (US)
Pages (from-to)3957-3969
Number of pages13
JournalJournal of Proteome Research
Volume14
Issue number9
DOIs
StatePublished - Jan 1 2015

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H3N2 Subtype Influenza A Virus
Glycosylation
Influenza A virus
Hemagglutinins
Viruses
Polysaccharides
Collectins
Pulmonary Surfactant-Associated Protein D
Glycopeptides
Immune system
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Membrane Glycoproteins
Zoonoses
Hong Kong
Pandemics
Mannose
Neutralizing Antibodies
Orthomyxoviridae
Oligosaccharides
Human Influenza

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

Glycosylation analysis of engineered H3N2 influenza A virus hemagglutinins with sequentially added historically relevant glycosylation sites. / An, Yanming; Mccullers, Jonathan; Alymova, Irina; Parsons, Lisa M.; Cipollo, John F.

In: Journal of Proteome Research, Vol. 14, No. 9, 01.01.2015, p. 3957-3969.

Research output: Contribution to journalArticle

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