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Volume 272, Number 43, Issue of October 24, 1997 pp. 27025-27034
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

The Glycosylation of Rat Intestinal Muc2 Mucin Varies between Rat Strains and the Small and Large Intestine
A STUDY OF O-LINKED OLIGOSACCHARIDES BY A MASS SPECTROMETRIC APPROACH

(Received for publication, July 8, 1997, and in revised form, August 18, 1997)

Niclas G. Karlsson Dagger , Annkatrin Herrmann , Hasse Karlsson Dagger , Malin E. V. Johansson Dagger , Ingemar Carlstedt and Gunnar C. Hansson Dagger

From the Dagger  Department of Medical Biochemistry, Göteborg University, Medicinaregatan 9A, 413 90 Gothenburg, Sweden and the  Department of Cell and Molecular Biology, Section for Molecular Pathogenesis, Lund University, P. O. Box 94, 221 00 Lund, Sweden

The large glycosylated domains obtained from the rat intestinal mucin Muc2 were isolated from the large and small intestine of the inbred rat strains GOT-W and GOT-BW. The expression of the rat Muc2 in the large intestine was confirmed immunochemically and by Northern blotting. Released oligosaccharides were structurally characterized by gas chromatography-mass spectrometry (neutral and sialylated species) or by tandem mass spectrometry (sulfated species), and a total of 63 structures was assigned. The large intestinal oligosaccharides were found to be identical between the strains, while the small intestinal glycosylation differed. Until now, detailed structural analysis of oligosaccharides isolated from a single mucin core or mucin domain with different origin have not been performed, and the information of different mucin glycoforms has been limited to immunochemistry. Blood group A-determinants (GalNAcalpha 1-3(Fucalpha 1-2)Galbeta 1-, and structures related to the blood group Sda/Cad-related epitope NeuAc/NeuGcalpha 1-3(GalNAcbeta 1-4)Galbeta 1-, were found in GOT-BW small intestine, and also in both large intestines. Blood group H-determinants and NeuAc/NeuGcalpha 1-3Galbeta 1- were found in all samples. Core 1 (Galbeta 1-3GalNAcalpha 1-), core 2 (Galbeta 1-3(GlcNAcbeta 1-6)GalNAcalpha 1-), core 3 (GlcNAcbeta 1-3GalNAcalpha 1-), and core 4 (GlcNAcbeta 1-3(GlcNAcbeta 1-6)GalNAcalpha 1- were also found in all the samples. The large intestine were enriched in sulfated oligosaccharides and the small intestine contained higher amounts of sialylated species. Sulfation were found exclusively on C-6 of GlcNAc.


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