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Biochemical and enzymatic characterization of blood group ABH and related histo-blood group glycosphingolipids in the epithelial cells of porcine small intestine.

Artikel i vetenskaplig tidskrift
Författare Annika E. Bäcker
Michael Breimer
B E Samuelsson
Jan Holgersson
Publicerad i Glycobiology
Volym 7
Nummer/häfte 7
Sidor 943-53
ISSN 0959-6658
Publiceringsår 1997
Publicerad vid Institutionen för de kirurgiska disciplinerna, Avdelningen för kirurgi
Sidor 943-53
Språk en
Länkar www.ncbi.nlm.nih.gov/entrez/query.f...
Ämnesord ABO Blood-Group System, chemistry, metabolism, Amino Acid Sequence, Animals, Carbohydrate Conformation, Chromatography, High Pressure Liquid, Epithelial Cells, enzymology, metabolism, Glycosphingolipids, chemistry, metabolism, Intestine, Small, enzymology, metabolism, Mass Spectrometry, Molecular Sequence Data, Swine
Ämneskategorier Klinisk medicin, Kirurgi

Sammanfattning

Non-acid glycosphingolipids were isolated from small intestinal epithelial cells of a single blood group A pig. One very predominant blood group compound was obtained chemically pure upon HPLC fractionation. It was characterized by mass spectrometry and 1H NMR spectroscopy to be the type 1 chain blood group A hexaglycosylceramide. Support for the presence of minute amounts of additional A glycolipids was obtained by mass spectrometry and immunostaining of TLC plates with anti-A antibodies specific for A type 2 chain, A type 3 and 4 chain, and the ALe(b) determinant. Among precursor chains, globoside (type 4) and lactotetraosylceramide (type 1) were immunologically identified, whereas no neolactotetraosylceramide (type 2) and gangliotetraosylceramide reactivities were detected. We addressed the question whether the predominant expression of type 1 chain based A glycolipids reflects a restricted glycolipid precursor chain specificity of the alpha 1-2 fucosyl- and/or the alpha 1-3 N-acetylgalactosaminyltransferases, or if the biosynthesis of the precursor chains themselves is regulated. All precursor core saccharides, lacto- (type 1), neolacto-(type 2), and gangliotetraosylceramide as well as globopentaosylceramide (type 4), could serve as acceptors for fucose in vitro when a crude microsomal fraction obtained from mechanically released, porcine intestinal epithelial cells was used as an enzyme source. Under the same conditions an N-acetylgalactosamine residue could be transferred to the blood group H structures based on these core saccharide chains. Lactotriaosylceramide, but not gangliotriaosylceramide, could serve as an acceptor for UDP-galactose. When the product was digested with beta-galactosidase (EC 3.2.1.23) from S.pneumoniae, under conditions where it specifically cleaves Gal beta 1-4 residues, approximately 40% of the radioactivity was cleaved off, indicating that a substantial amount of neolactotetraosylceramide was made in vitro, as opposed to the predominance of lactotetraosylceramide-based structures found in vivo.

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