A Sialylated Glycan Microarray Reveals Novel Interactions of Modified Sialic Acids with Proteins and Viruses*

• Type of Material: Article
• Authors: Xuezheng Song, Emory University; Hai Yu, University of California Davis; Xi Chen, Emory University; Yi Lasanajak, Emory University; Mary M. Tappert, University of Oklahoma; Gillian M. Air, University of Oklahoma; Vinod K. Tiwari, University of California Davis; Hongzhi Cao, University of California Davis; Harshal A. Chokhawala, University of California Davis; Haojie Zheng, University of California Davis; Richard D. Cummings, Emory University; David Smith, Emory University
• Language: English
• Date: 2011-09-09
• Publisher: American Society for Biochemistry and Molecular Biology
• Publication Version: Final Published Version
• Copyright Statement: © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
• Final Published Version (URL): http://www.jbc.org/content/286/36/31610
• Title of Journal or Parent Work: Journal of Biological Chemistry
• ISSN: 0021-9258
• Volume: 286
• Issue: 36
• Start Page: 31610
• End Page: 31622
• Grant/Funding Information: This work was also supported by Defense Advanced Research Projects Agency Grant HR0011-10-00 (to R. D. C.) and Oklahoma Center for the Advancement of Science and Technology Grant HR09-001 (to G. M. A.).; This work was supported, in whole or in part, by National Institutes of Health (NIH) Grant RO1GM085448 (to D. F. S.), a bridging grant from the Consortium for Functional Glycomics under NIGMS, NIH Grant GM62116 (to R. D. C.), and NIH Grant GM076360 (to X. C.).
• Keywords: Functional Glycomics; Sialic Acid; Lectin; Protein-Carbohydrate Interactions; Glycomics; Influenza Virus; Microarray
• Research Categories: Chemistry, Biochemistry

Abstract

Many glycan-binding proteins in animals and pathogens recognize sialic acid or its modified forms, but their molecular recognition is poorly understood. Here we describe studies on sialic acid recognition using a novel sialylated glycan microarray containing modified sialic acids presented on different glycan backbones. Glycans terminating in β-linked galactose at the non-reducing end and with an alkylamine-containing fluorophore at the reducing end were sialylated by a one-pot three-enzyme system to generate α2–3- and α2–6-linked sialyl glycans with 16 modified sialic acids. The resulting 77 sialyl glycans were purified and quantified, characterized by mass spectrometry, covalently printed on activated slides, and interrogated with a number of key sialic acid-binding proteins and viruses. Sialic acid recognition by the sialic acid-binding lectins Sambucus nigra agglutinin and Maackia amurensis lectin-I, which are routinely used for detecting α2–6- and α2–3-linked sialic acids, are affected by sialic acid modifications, and both lectins bind glycans terminating with 2-keto-3-deoxy-d-glycero-d-galactonononic acid (Kdn) and Kdn derivatives stronger than the derivatives of more common N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc). Three human parainfluenza viruses bind to glycans terminating with Neu5Ac or Neu5Gc and some of their derivatives but not to Kdn and its derivatives. Influenza A virus also does not bind glycans terminating in Kdn or Kdn derivatives. An especially novel aspect of human influenza A virus binding is its ability to equivalently recognize glycans terminated with either α2–6-linked Neu5Ac9Lt or α2–6-linked Neu5Ac. Our results demonstrate the utility of this sialylated glycan microarray to investigate the biological importance of modified sialic acids in protein-glycan interactions.