Glycobiology, 2001, Vol. 11, No. 5 373-384
© 2001 Oxford University Press
Characterization of N-glycans from mouse brain neural cell adhesion molecule
3Zentrum für Molekulare Neurobiologie, Universität Hamburg, D-20246 Hamburg, Germany, 4Institute of Biochemistry, University of Giessen, D-35392 Giessen, Germany, 5Institute for Molecular Biology and Biophysics, ETH Hönggerberg, CH-8093 Zürich, Switzerland, 6Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, D-30625 Hannover, Germany, and 7Division of Immunochemistry, Research Center Borstel, D-23845 Borstel, Germany
The N-glycosylation pattern of the neural cell adhesion molecule (NCAM), isolated from brains of newborn mice, has been analyzed. Following digestion with trypsin, generated glycopeptides were fractionated by serial immunoaffinity chromatography using immobilized monoclonal antibodies specifically recognizing polysialic acid (PSA) units or the HNK1-carbohydrate epitope. Subsequent analyses of the resulting (glyco)peptides by Edman degradation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) revealed polysialylated glycans to be exclusively linked to glycosylation sites 5 (Asn431) and 6 (Asn460), whereas glycans carrying the HNK1-epitope could be assigned to sites 2 (Asn297), 5, 6, and, to a lesser extent, site 3 (Asn329). PSA-, HNK1-, and non-PSA/HNK1-glycan fractions were characterized by carbohydrate constituent and methylation analyses as well as MALDI-TOF-MS in conjunction with chromatographic fractionation techniques. The results revealed that the core structures of PSA-glycans represented predominantly fucosylated, partially sulfated 2,6-branched isomers of triantennary as well as tetraantennary complex-type glycans, whereas carbohydrate chains bearing the HNK1-epitope were dominated by diantennary species carrying in part bisecting GlcNAc residues. Non-PSA/HNK1-glycans exhibited a highly heterogeneous pattern of partially truncated, mostly diantennary structures being characterized by the presence of additional fucose, bisecting GlcNAc and/or sulfate residues. In conclusion, our results revealed that the glycosylation pattern of murine NCAM displays high structural and regional selectivity, which might play an important role in controlling the biological activities of this molecule.
1 Present address: bai GmbH, D-64686 Lautertal, Germany.
2 To whom correspondence should be addressed at: Biochemisches Institut am Klinikum der Universität, Friedrichstrasse 24, D-35392 Giessen, Germany.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  D. Anzai, Y. Tonoyama, A. Ikeda, T. Kawasaki, and S. Oka Regulated expression of the HNK-1 carbohydrate is essential for medaka (Oryzias latipes) embryogenesis Glycobiology, August 1, 2009; 19(8): 868 - 878. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Yoshihara, K. Sugihara, Y. Kizuka, S. Oka, and M. Asano Learning/Memory Impairment and Reduced Expression of the HNK-1 Carbohydrate in {beta}4-Galactosyltransferase-II-deficient Mice J. Biol. Chem., May 1, 2009; 284(18): 12550 - 12561. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Morita, Y. Kizuka, S. Kakuda, and S. Oka Expression and Function of the HNK-1 Carbohydrate J. Biochem., June 1, 2008; 143(6): 719 - 724. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kizuka, K. Kobayashi, S. Kakuda, Y. Nakajima, S. Itoh, N. Kawasaki, and S. Oka Laminin-1 is a novel carrier glycoprotein for the nonsulfated HNK-1 epitope in mouse kidney Glycobiology, April 1, 2008; 18(4): 331 - 338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Oltmann-Norden, S. P. Galuska, H. Hildebrandt, R. Geyer, R. Gerardy-Schahn, H. Geyer, and M. Muhlenhoff Impact of the Polysialyltransferases ST8SiaII and ST8SiaIV on Polysialic Acid Synthesis during Postnatal Mouse Brain Development J. Biol. Chem., January 18, 2008; 283(3): 1463 - 1471. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Galuska, R. Geyer, R. Gerardy-Schahn, M. Muhlenhoff, and H. Geyer Enzyme-dependent Variations in the Polysialylation of the Neural Cell Adhesion Molecule (NCAM) in Vivo J. Biol. Chem., January 4, 2008; 283(1): 17 - 28. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Sjostrand, J. Carlsson, G. Paratcha, B. Persson, and C. F. Ibanez Disruption of the GDNF Binding Site in NCAM Dissociates Ligand Binding and Homophilic Cell Adhesion J. Biol. Chem., April 27, 2007; 282(17): 12734 - 12740. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Sandra, P. Dolashka-Angelova, B. Devreese, and J. Van Beeumen New insights in Rapana venosa hemocyanin N-glycosylation resulting from on-line mass spectrometric analyses Glycobiology, February 1, 2007; 17(2): 141 - 156. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Galuska, I. Oltmann-Norden, H. Geyer, B. Weinhold, K. Kuchelmeister, H. Hildebrandt, R. Gerardy-Schahn, R. Geyer, and M. Muhlenhoff Polysialic Acid Profiles of Mice Expressing Variant Allelic Combinations of the Polysialyltransferases ST8SiaII and ST8SiaIV J. Biol. Chem., October 20, 2006; 281(42): 31605 - 31615. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kizuka, T. Matsui, H. Takematsu, Y. Kozutsumi, T. Kawasaki, and S. Oka Physical and Functional Association of Glucuronyltransferases and Sulfotransferase Involved in HNK-1 Biosynthesis J. Biol. Chem., May 12, 2006; 281(19): 13644 - 13651. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Nakata and F. A. Troy II Degree of Polymerization (DP) of Polysialic Acid (PolySia) on Neural Cell Adhesion Molecules (N-CAMs): DEVELOPMENT AND APPLICATION OF A NEW STRATEGY TO ACCURATELY DETERMINE THE DP OF polySIA CHAINS ON N-CAMS J. Biol. Chem., November 18, 2005; 280(46): 38305 - 38316. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Yagi, N. Takahashi, Y. Yamaguchi, N. Kimura, K. Uchimura, R. Kannagi, and K. Kato Development of structural analysis of sulfated N-glycans by multidimensional high performance liquid chromatography mapping methods Glycobiology, October 1, 2005; 15(10): 1051 - 1060. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Tagawa, Y. Kizuka, T. Ikeda, S. Itoh, N. Kawasaki, H. Kurihara, M. L. Onozato, A. Tojo, T. Sakai, T. Kawasaki, et al. A Non-sulfated Form of the HNK-1 Carbohydrate Is Expressed in Mouse Kidney J. Biol. Chem., June 24, 2005; 280(25): 23876 - 23883. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. E. Close, S. S. Mendiratta, K. M. Geiger, L. J. Broom, L.-L. Ho, and K. J. Colley The Minimal Structural Domains Required for Neural Cell Adhesion Molecule Polysialylation by PST/ST8Sia IV and STX/ST8Sia II J. Biol. Chem., August 15, 2003; 278(33): 30796 - 30805. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. L. Poongodi, N. Suresh, S. C. B. Gopinath, T. Chang, S. Inoue, and Y. Inoue Dynamic Change of Neural Cell Adhesion Molecule Polysialylation on Human Neuroblastoma (IMR-32) and Rat Pheochromocytoma (PC-12) Cells during Growth and Differentiation J. Biol. Chem., July 26, 2002; 277(31): 28200 - 28211. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Ong, M. Suzuki, F. Belot, J.-C. Yeh, I. Franceschini, K. Angata, O. Hindsgaul, and M. Fukuda Biosynthesis of HNK-1 Glycans on O-Linked Oligosaccharides Attached to the Neural Cell Adhesion Molecule (NCAM). THE REQUIREMENT FOR CORE 2 beta 1,6-N-ACETYLGLUCOSAMINYLTRANSFERASE AND THE MUSCLE-SPECIFIC DOMAIN IN NCAM J. Biol. Chem., May 10, 2002; 277(20): 18182 - 18190. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. von der Ohe, S. F. Wheeler, M. Wuhrer, D. J. Harvey, S. Liedtke, M. Muhlenhoff, R. Gerardy-Schahn, H. Geyer, R. A. Dwek, R. Geyer, et al. Localization and characterization of polysialic acid-containing N-linked glycans from bovine NCAM Glycobiology, January 1, 2002; 12(1): 47 - 63. [Abstract] [Full Text] [PDF]
|
|