Shotgun glycomics of pig lung identifies natural endogenous receptors for influenza viruses

• Authors: Lauren Byrd-Leotis, Emory University; Renpeng Liu, Emory University; Konrad C. Bradley, Emory University; Yi Lasanajak, Emory University; Sandra F. Cummings, Emory University; Xuezheng Song, Emory University; Jamie Heimburg-Molinaro, Emory University; Summer Galloway, Emory University; Marie R. Culhane, University of Minnesota; David Smith, Emory University; David Steinhauer, Emory University; Richard Cummings, Emory University
• Journal Title: Proceedings of the National Academy of Sciences
• Volume: Volume 111, Number 22
• Publisher: National Academy of Sciences | 2014-06-03, Pages E2241-E2250
• Type of Work: Article | Final Publisher PDF
• Publisher DOI: http://doi.org/10.1073/pnas.1323162111
• Subjects: Biology, Microbiology; Chemistry, Biochemistry
• Keywords: Science & Technology; Multidisciplinary Sciences; Science & Technology - Other Topics; receptor binding; hemagglutinin; virus attachment; RICINUS-COMMUNIS AGGLUTININ; GLYCAN-BINDING-PROTEINS; O-LINKED GLYCANS; L BARK LECTIN; SIALIC-ACID; A VIRUSES; STRUCTURAL DETERMINANTS; MICROARRAY STRATEGY; PANDEMIC INFLUENZA; MASS-SPECTROMETRY

Abstract

Influenza viruses bind to host cell surface glycans containing terminal sialic acids, but as studies on influenza binding become more sophisticated, it is becoming evident that although sialic acid may be necessary, it is not sufficient for productive binding. To better define endogenous glycans that serve as viral receptors, we have explored glycan recognition in the pig lung, because influenza is broadly disseminated in swine, and swine have been postulated as an intermediary host for the emergence of pandemic strains. For these studies, we used the technology of "shotgun glycomics" to identify natural receptor glycans. The total released N- and O-glycans from pig lung glycoproteins and glycolipid-derived glycans were fluorescently tagged and separated by multidimensional HPLC, and individual glycans were covalently printed to generate pig lung shotgun glycan microarrays. All viruses tested interacted with one or more sialylated N-glycans but not O-glycans or glycolipid-derived glycans, and each virus demonstrated novel and unexpected differences in endogenous N-glycan recognition. The results illustrate the repertoire of specific, endogenous N-glycans of pig lung glycoproteins for virus recognition and offer a new direction for studying endogenous glycan functions in viral pathogenesis.