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Article
NMR Detection and Characterization of Sialylated Glycoproteins and Cell Surface Polysaccharides
Journal of Biomolecular NMR (2011)
  • Adam Barb, University of Georgia
  • Darón I. Freedberg
  • Marcos D. Battistel
  • James H. Prestegard
Abstract
Few solution NMR pulse sequences exist that are explicitly designed to characterize carbohydrates (glycans). This is despite the essential role carbohydrate motifs play in cell–cell communication, microbial pathogenesis, autoimmune disease progression and cancer metastasis, and despite that fact that glycans, often shed to extra-cellular fluids, can be diagnostic of disease. Here we present a suite of two dimensional coherence experiments to measure three different correlations (H3–C2, H3–C1, and C1–C2) on sialic acids, a group of nine-carbon carbohydrates found on eukaryotic cell surfaces that often play a key role in disease processes. The chemical shifts of the H3, C2, and C1 nuclei of sialic acids are sensitive to carbohydrate linkage, linkage conformation, and ionization state of the C1 carboxylate. The experiments reported include rigorous filter elements to enable detection and characterization of isotopically labeled sialic acids with high sensitivity in living cells and crude isolates with minimal interference from unwanted signals arising from the ~1% 13C-natural abundance of cellular metabolites. Application is illustrated with detection of sialic acids on living cells, in unpurified mixtures, and at the terminus of the N-glycan on the 55 kDa immunoglobulin G Fc.
Keywords
  • N-acetylneuraminic acid,
  • Sialic acid,
  • Carbohydrate,
  • Pulse sequence,
  • In vivo NMR,
  • Isotopic labeling
Publication Date
September, 2011
Publisher Statement
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.
Citation Information
Adam Barb, Darón I. Freedberg, Marcos D. Battistel and James H. Prestegard. "NMR Detection and Characterization of Sialylated Glycoproteins and Cell Surface Polysaccharides" Journal of Biomolecular NMR Vol. 51 Iss. 1-2 (2011)
Available at: http://works.bepress.com/adam_barb/1/