University of Maryland Baltimore County

Instructor: C. Allen Bush - Room 208 Chemistry ext. 5-2506

and Prof. Lai-Xi Wang, IHV, Univ. of Md. Medical School..

Text: "Functional and Molecular Glycobiology"

by S.A. Brooks, M.V.Dwek, U. Schumacher BIOS Scientific Publishers, Ltd. Oxford, 2002

• 1. Basic chemistry of carbohydrates.

  • Monosaccharides, stereochemistry, chemical functionality.
  • Glycosides, disaccharide linkage, oligosaccharides, polysaccharides
• 2. Glycoproteins

  • Constituent monosaccharides, Covalent glycopeptide linkages, Microheterogenity
  • Asparagine N-linked glycopeptides. Glycosylation of Recombinant glycoproteins.
  • High Mannose chains - ovalbumin IgM, Complex antenna chains - sialic acids.
  • Glycosidase digestion of N-linked glycopeptides
  • exo- and endoglycosidases, endo H, endo F, endo b-galactosidase, peptide N-glycanase
  • Mucin glycoproteins - O-glycosidic linkage -
  • antifreeze glycoproteins, ovarian cyst mucins, blood group substances.
• 3. Glycosaminoglycans and Proteoglycans - Chondroitin, hyaluronic acids, Heparin, dermatan. Cartilage, basement membrane and cell surface proteoglycans.
• 4. Plasma Membranes - carbohydrates at the outer surface of cells. Membrane glycoproteins. Red cell glycophorin, band 3.
• 5. Glycolipids. sphingosine, ceramides

  • gangliosides, globosides, human milk oligosaccharides
  • Tumor antigens. Lewis blood groups. Sialyl Lewis^X type, Embryonic antigens
  • Role of Lewis oligosaccharides in inflammation and tumor metastasis.
  • Glyco-phosphatidyl Inositol anchors. Membrane glycoprotein anchors compared with peptide anchors. Trypanosome variant surface glycoprotein glycan structure.
• 6. Biosynthesis of Complex Carbohydrates

  • Biosynthesis of N-linked glycoproteins. endoplasmic reticulum, golgi, lipid-linked oligosaccharide, dolichol.
  • Oligosachryl transferase
  • Processing of the oligosaccharide, resynthesis.
  • Cellular targeting of the glycoprotein.
  • Role of glycosylation in protein folding.
  • Biosynthesis of O-linked glycoproteins. Mucins.
• 7. Chemical and enzymatic synthesis of complex oligosaccharides and glycopeptides.

  • Chemical reactions for glycosylation. Protecting groups. Regioselectivity and stereoselectivity.
  • Enzymatic glycosylation, mechanisms of glcosyltransferases. Transglycosylation by glycosidases
• 8. Bacterial Cell Surface Carbohydrates

  • Cell wall peptidoglycan, cell wall polysaccharides, capsules
  • extracellular polysaccharides, lipopolysaccharides, endotoxin, lipid A, O-antigens
  • Genes in the synthesis of bacterial polysaccharides: glycosyl transferases, membrane transport proteins, polymerases.
  • Bacterial adhesion in pathogenesis. Bacterial polysaccharide vaccines.
• 9. Protein-Carbohydrate interaction -

  • Lectins decode the information stored in complex carbohydrates
  • Plant lectins, Con-A, ricin, blood typing, mitogenic stimulation
  • Bacterial adhesion, polysaccharides and bacterial lectins.
  • S- and C-type animal lectins. Soluble lectins and tissue-specific differentiation.
  • Selectins and inflammatory response.
  • Humoral and cell associated lectins from invertebrates.
• 10. Structural Analysis of complex carbohydrates

  • Chromatography. HPLC, GC analysis of sugar composition.
  • Oligosaccharide fractionation by HPLC
  • Methylation analysis, Smith degradation.
  • Mass spectrometry for structure determination.
  • NMR analysis by ¹H and ¹³C spectroscopy.
  • Conformation and Dynamics of Oligosaccharides
  • x-ray crystallography, NMR spectroscopy, conformational modeling