Structure of the asparagine-linked sugar chains of porcine kidney and human urine cerebroside sulfate activator protein.

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  • Additional Information
    • Source:
      Publisher: Wiley Country of Publication: England NLM ID: 9504818 Publication Model: Print Cited Medium: Print ISSN: 1076-5174 (Print) Linking ISSN: 10765174 NLM ISO Abbreviation: J Mass Spectrom Subsets: MEDLINE
    • Publication Information:
      Original Publication: Chichester, UK : Wiley, c1995-
    • Subject Terms:
      Carbohydrate Conformation*; Asparagine/*chemistry ; Glycoproteins/*chemistry ; Kidney/*chemistry; Animals ; Chromatography, High Pressure Liquid ; Glycoproteins/urine ; Humans ; Molecular Structure ; Saposins ; Spectrometry, Mass, Electrospray Ionization ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Sphingolipid Activator Proteins ; Swine
    • Abstract:
      The specific sugar residues and their linkages in the oligosaccharides from pig kidney and human urine cerebroside sulfate activator proteins (saposin B), although previously hypothesized, have been unambiguously characterized. Exhaustive sequential exoglycosidase digestion of the trimethyl-p-aminophenyl derivatives, followed by either matrix-assisted laser desorption/ionization and/or mass spectrometry, was used to define the residues and their linkages. The oligosaccharides were enzymatically released from the proteins by treatment with peptidyl-N-glycosidase F and separated from the proteins by reversed-phase high-performance liquid chromatography (HPLC). Reducing termini were converted to the trimethyl-p-aminophenyl derivative and the samples were further purified by normal-phase HPLC. The derivatized carbohydrates were then treated sequentially with a series of exoglycosidases of defined specificity, and the products of each digestion were examined by mass spectrometry. The pentasaccharides from pig kidney and human urine protein were shown to be of the asparagine-linked complex type composed of mannose-alpha 1-6-mannose-beta 1-4-N-acetylglucosamine-N-acetylglucosamine(alpha 1-6-fucose). This highly degraded structure probably represents the final product of intra-lysosomal exoglycosidase digestion. Oligosaccharide sequencing by specific exoglycosidase degradation coupled with mass spectrometry is more rapid than conventional oligosaccharide sequencing. The procedures developed will be useful for sequencing other oligosaccharides including those from other members of the lipid-binding protein class to which cerebroside sulfate activator belongs. (c) 2000 John Wiley & Sons, Ltd.
    • Grant Information:
      NS31271 United States NS NINDS NIH HHS
    • Accession Number:
      0 (Glycoproteins)
      0 (PSAP protein, human)
      0 (Saposins)
      0 (Sphingolipid Activator Proteins)
      7006-34-0 (Asparagine)
    • Publication Date:
      Date Created: 20010217 Date Completed: 20010405 Latest Revision: 20071114
    • Publication Date:
      20240829
    • Accession Number:
      10.1002/1096-9888(200012)35:12<1416::AID-JMS75>3.0.CO;2-K
    • Accession Number:
      11180632