Shotgun Glycomics: A Microarray Strategy for Functional Glycomics

• Type of Material: Article
• Authors: Xuezheng Song, Emory University; Yi Lasanajak, Emory University; Baoyun Xia, Emory University; Jamie Heimburg-Molinaro, Emory University; Jeanne M. Rhea, Emory University; Hong Ju, Emory University; Chunmei Zhao, Emory University; Ross J Molinaro, Emory University; Richard D. Cummings, Emory University; David Smith, Emory University
• Language: English
• Date: 2011-01
• Publisher: Nature Research (part of Springer Nature)
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2011 Nature America, Inc. All rights reserved.
• Final Published Version (URL): http://www.nature.com/nmeth/journal/v8/n1/full/nmeth.1540.html
• Title of Journal or Parent Work: Nature Methods
• ISSN: 1548-7091
• Volume: 8
• Issue: 1
• Start Page: 85
• End Page: 90
• Grant/Funding Information: This work was supported by in part by a Bridge Grant to R.D.C from the Consortium for Functional Glycomics from the National Institute of General Medical Sciences (Grant GM62116) and a EUREKA Grant (GM08544) to D.F.S. from the National Institute of General Medical Sciences.; National Institute of General Medical Sciences : NIGMS
• Keywords: glycosphingolipids; immobilization; fluorescent labeling; functional glycomics; glycan array
• Research Categories: Chemistry, Biochemistry

Abstract

Major challenges of glycomics are to characterize a glycome and identify functional glycans as ligands for glycan-binding proteins (GBPs). To address these issues we have developed a general strategy termed shotgun glycomics. We focus on glycosphingolipids (GSLs), a challenging class of glycoconjugates recognized by toxins, antibodies, and GBPs. We derivatized GSLs extracted from cells with a heterobifunctional fluorescent tag suitable for covalent immobilization. Fluorescent GSLs were separated by multidimensional chromatography, quantified, and coupled to glass slides to create GSL shotgun microarrays. The microarrays were interrogated with cholera toxin, antibodies, and sera from patients with Lyme disease to identify biologically relevant GSLs that were subsequently characterized by mass spectrometry. Shotgun glycomics incorporating GSLs and potentially glycoprotein-derived glycans provides an approach to accessing the complex glycomes of animal cells and offers a strategy for focusing structural analyses on functionally significant glycans.