Abstract
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 (GalNAcalpha1-3(Fucalpha1-2)Galbeta1-, and structures related to the blood group Sda/Cad-related epitope NeuAc/NeuGcalpha1-3(GalNAcbeta1-4)Galbeta1-, were found in GOT-BW small intestine, and also in both large intestines. Blood group H-determinants and NeuAc/NeuGcalpha1-3Galbeta1- were found in all samples. Core 1 (Galbeta1-3GalNAcalpha1-), core 2 (Galbeta1-3(GlcNAcbeta1-6)GalNAcalpha1-), core 3 (GlcNAcbeta1-3GalNAcalpha1-), and core 4 (GlcNAcbeta1-3(GlcNAcbeta1-6)GalNAcalpha1- 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.
Highlights
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
To further explore the glycosylation of a mucin domain from Muc2, we studied the glycosylation of the small intestinal glycopeptide A (gpA) from a black-and-white inbred rat strain (GOT-BW)
1, starting dilution 1:50, GSL standard H5–1. f MAb DAKO A583, starting dilution 1:25, GSL standard H5–2. g Values given are reactivity of 400 ng of HF-deglycosylated gpA with polyclonal antibody raised against HF-deglycosylated gpA from GOT-W small intestine (␣-gpA, PH497)
Summary
We have previously reported a detailed structural characterization of the oligosaccharides from gpA and gpB from an inbred white rat strain (GOT-W), known to express H-determinants but not A-determinants in the small intestine [2, 8]. To further explore the glycosylation of a mucin domain from Muc, we studied the glycosylation of the small intestinal gpA from a black-and-white inbred rat strain (GOT-BW). This strain has been shown to express blood group A positive glycosphingolipids in the small intestine [13]. Detailed qualitative and quantitative information from GC-MS, FAB-MS, and MS/MS revealed that the Muc2-derived mucin domain gpA existed as different glycoforms in the small and large intestine. It was further concluded that the small intestinal glycosylation of this domain differed between the two inbred rat strains
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
