Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites

• DOI: 10.1021/acs.jproteome.5b00416
• Publication Date: 2015-07-23T02:07:47Z
• Authors: Yanming An; Jonathan A. McCul...; Irina Alymova; Lisa M. Parsons; John F. Cipollo
• Keywords: Collectin; Glycomics; N-linked glycosylation

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