caa a22 4500 469115092 CHVBK 20180323133122.0 cr unu---uuuuu 170328e19921201xx s 000 0 eng 10.1007/BF00731089 doi (NATIONALLICENCE)springer-10.1007/BF00731089 Use of large-scale hydrazinolysis in the preparation of N -linked oligosaccharide libraries: application to brain tissue [Elektronische Daten] [D. Wing, T. Rademacher, M. Field, R. Dwek, B. Schmitz, G. Thor, M. Schachner] In this report, we describe the preparation of a library ofN-linked glycans from whole murine brain obtained by the large-scale hydrazinolysis of an acetone powder of the tissue followed by chromatographic procedures. 84% of the characterized oligosaccharides were found to be anionic, the remainder neutral. The anionic species were successively neutralized by neuraminidase (29%), aq. hydrofluoric acid (30%), and methanolysis (26%), indicating that approximately equal portions were sensitive to desialylation, dephosphorylation and desulfation, respectively. The presence of the sulfated fraction was confirmed by direct35SO4 metabolic labelling. A residual partially characterized fraction was found to be anionic through possession of carboxylic acid groups, unrelated to sialic acid. The purified oligosaccharides, in the absence of their original protein conjugates, were shown to retain those immunological characteristics essential for recognition by a specific monoclonal antibody, LS (412), that is known to recognize a carbohydrate epitope present on a number of neural adhesion molecules and functional in neural cell adhesion. These properties confirm the viability of scaling up the size of the hydrazinolysis procedure and adapting it to whole tissue for the production of glycan libraries and for the probing of structures of interest. Chapman & Hall, 1992 N -linked nationallicence oligosaccharide library nationallicence L2/HNK-1 epitope nationallicence anionicity nationallicence murine brain nationallicence large scale hydrazinolysis nationallicence ConA : concanavalin A nationallicence ELISA : enzyme-linked immunosorbent assay nationallicence Fuc : fucose nationallicence Gal : galactose nationallicence GalNAc : N-acetylgalactosamine nationallicence GlcNAc : N-acetylglucosamine nationallicence g.u. : glucose units nationallicence HRP : horseradish peroxidase nationallicence HVE : high voltage electrophoresis nationallicence Man : mannose nationallicence MS : mass spectrometry nationallicence N-CAM : neural cell adhesion molecule nationallicence Wing D. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut Rademacher T. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut Field M. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut Dwek R. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut Schmitz B. Department of Neurobiology, ETH-Hönggerberg, HPM, 8093, Zürich, Switzerland aut Thor G. Department of Neurobiology, ETH-Hönggerberg, HPM, 8093, Zürich, Switzerland aut Schachner M. Department of Neurobiology, ETH-Hönggerberg, HPM, 8093, Zürich, Switzerland aut Glycoconjugate Journal Kluwer Academic Publishers 9/6(1992-12-01), 293-301 0282-0080 9:6<293 1992 9 10719 https://doi.org/10.1007/BF00731089 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00731089 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wing D. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut NATIONALLICENCE 700 1- Rademacher T. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut NATIONALLICENCE 700 1- Field M. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut NATIONALLICENCE 700 1- Dwek R. Department of Biochemistry, Glycobiology Institute, South Parks Road, OX1 3QU, Oxford, UK aut NATIONALLICENCE 700 1- Schmitz B. Department of Neurobiology, ETH-Hönggerberg, HPM, 8093, Zürich, Switzerland aut NATIONALLICENCE 700 1- Thor G. Department of Neurobiology, ETH-Hönggerberg, HPM, 8093, Zürich, Switzerland aut NATIONALLICENCE 700 1- Schachner M. Department of Neurobiology, ETH-Hönggerberg, HPM, 8093, Zürich, Switzerland aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 9/6(1992-12-01), 293-301 0282-0080 9:6<293 1992 9 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer