caa a22 4500 445325577 CHVBK 20180317142719.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s10858-011-9550-0 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9550-0 NMR detection and characterization of sialylated glycoproteins and cell surface polysaccharides [Elektronische Daten] [Adam Barb, Darón Freedberg, Marcos Battistel, James Prestegard] 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 55kDa immunoglobulin G Fc. Springer Science+Business Media B.V., 2011 N -acetylneuraminic acid nationallicence Sialic acid nationallicence Carbohydrate nationallicence Pulse sequence nationallicence In vivo NMR nationallicence Isotopic labeling nationallicence Barb Adam Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, 30602, Athens, GA, USA aut Freedberg Darón Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, Food and Drug Administration, Building 29, Room 115, 1401 Rockville Pike, HFM-428, 20852, Rockville, MD, USA aut Battistel Marcos Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, Food and Drug Administration, Building 29, Room 115, 1401 Rockville Pike, HFM-428, 20852, Rockville, MD, USA aut Prestegard James Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, 30602, Athens, GA, USA aut Journal of Biomolecular NMR Springer Netherlands 51/1-2(2011-09-01), 163-171 0925-2738 51:1-2<163 2011 51 10858 https://doi.org/10.1007/s10858-011-9550-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9550-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Barb Adam Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, 30602, Athens, GA, USA aut NATIONALLICENCE 700 1- Freedberg Darón Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, Food and Drug Administration, Building 29, Room 115, 1401 Rockville Pike, HFM-428, 20852, Rockville, MD, USA aut NATIONALLICENCE 700 1- Battistel Marcos Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, Food and Drug Administration, Building 29, Room 115, 1401 Rockville Pike, HFM-428, 20852, Rockville, MD, USA aut NATIONALLICENCE 700 1- Prestegard James Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, 30602, Athens, GA, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/1-2(2011-09-01), 163-171 0925-2738 51:1-2<163 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer