1. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions

2. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions

3. The mammalian cell surface ?
Martinez-Palomo, A., et al. Cancer Res. 29, , 1969
The “glycocalyx” revealed (glycans stained with ruthinium red)
intracellular
fibroblast
extracellular
lipid bilayer
intracellular
fibroblast
extracellular
lipid bilayer
wiki/Plasma_membrane
Krijnse Locker J. & Schmid S.L. PLoS Biol 11, e , 2013
“Every eukaryotic cell is covered with a dense and complex array of glycans.” … in fact…
“Evolution has failed to generate a [single] living cell devoid of surface glycosylation” - A. Varki

4. What if a MAJOR cell component was invisible by standard microscopy?
Light microscopy micrograph of Cryptococcus neoformans capsule delineated by India ink. The inner circle represents the fungal cell, with the wide outer circle being the capsule.
Steenbergen et al. (2003) Microbes and Infection 5:667
Light microscopy micrograph of Cryptococcus neoformans
Steenbergen et al. (2003) Microbes and Infection 5:667
Scanning electron microscopy of C. neoformans yeast cells.
Van Duin et al. (2004) Antimicrobial Agents and Chemotherapy 48:2014

5. Each prokaryote carries a surface glycan coat
Electron microscopic thin section of Escherichia coli K1
Amako et al. (1988) J Bacteriol 170:4960
Electron microscopic thin section of Klebsiella pneumoniae
Amako et al. (1988) J Bacteriol 170:4960

6. CD59, a complement defense glycoprotein
Proteins typically fold onto themselves, whereas glycans spread out in space
3 kDa N-linked glycan
20 kDa protein
CD59, a complement defense glycoprotein
1 kDa O-linked glycan
1.5 kDa GPI anchor
plasma membrane

7. Glycoprotein
Glycolipid
plasma membrane

8. Tropical Forest Canopy
Ajit Varki, “Perspectives on the Future of Glycobiology,” Athens, GA, 14-Mar-2011

9. Tropical Forest Canopy
Cohen & Varki (2010) OMICS 4:455

10. The Cell Surface textbook image real image lipid bilayer fibroblast
intracellular
Martinez-Palomo, A., et al. Cancer Res. 29, , 1969

11. Glycan-mediated molecular recognition at the cell surface
Lectin
Glycan
Sharon and Lis (1993) Scientific American

12. Life cycle of the influenza virus - the role of glycans

13. Influenza jumps from animals to humans via a slight change in glycan binding specificity
Stevens, et al. (2006) Nat Rev Microbiol. 4:857-64

14. Influenza jumps from animals to humans via a slight change in glycan binding specificity
Stevens, et al. (2006) Nat Rev Microbiol. 4:857-64

15. Inflammation: How we fight bacterial infection

16. You fight bacterial infection using glycans. This child could not …
Marquardt, et al. (1999) J. Pediatr. 134, 681

17. …until he was fed FUCOSE (a sugar), after which…
Marquardt, et al. (1999) Blood 94, 3976
“…neutrophil levels returned to the normal range .... there were no further infections”

18. Glycan-mediated molecular recognition at the cell surface
Lectin
Glycan
Sharon and Lis (1993) Scientific American

19. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions

20. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Just nine monosaccharide saccharide building blocks are responsible for building nearly all mammalian glycans.
All are six-membered rings (5 carbons, 1 oxygen) varying in the substituents at each carbon (mostly hydroxyls).
Seven are closely structurally related to glucose.

21. Seven sugars and their structural relationship to glucose
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition

22. Two important terminal eukaryotic sugars
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition

23. Monosaccharides equilibrate between open chain and ring forms
Monosaccharides equilibrate between open chain and ring forms. For each monosaccharide, two ring forms (anomeric configurations) are possible, α and β

24. When a glycosidic bond is formed, the anomeric configuration is “locked”

25. Nomenclature:
-Name the non-reducing (left-most) sugar (Gal) -Name the anomeric configuration (β) -Name the anomeric carbon number (1) -Name the substituted carbon number (4) -Name the substituted sugar (Glc)
RESULT: Gal β1-4 Glc

26. Nomenclature: Branches are placed in parentheses
NeuAc α2-3 Gal β1-4 (Fuc α1-3) GlcNAc

27. NeuAc α2-3 Gal β1-4 (Fuc α1-3) GlcNAc

28. Molecular diversity of glycans
Polypeptides
Glycans
Building blocks
amino acids
monosaccharides
Number of different monomers
20 common
9 common
Linkage sites per monomer 1
3-4
Possible linkage configurations 2
Possible homodimer structures
6-8
Linkage modes
linear
linear or branched

29. Three different amino acids (Ala, Ser, Tyr) – 6 structures Ala-Ser-Tyr   
Ala-Ser-Tyr
Ala-Tyr-Ser
Ser-Ala-Tyr
Ser-Tyr-Ala
Tyr-Ala-Ser
Tyr-Ser-Ala
Three different sugars (Glc, Gal, Man) – 1,056 structures …

30. Man a1-4 Gal a1-3 Glc b1 Man a1-6 Gal a1-3 Glc b1
 Gal a1-2 Glc a1-2 Man a1-
Gal a1-3 Glc a1-6 Man b1-
Gal a1-4 Man b1-4 Glc a1-
Man a1-6 Glc a1-3 Gal a1-
Man b1-2 Glc b1-2 Gal b1-
Man b1-3 Gal b1-6 Glc a1-
Glc b1-4 Man a1-4 Gal a1-
Glc b1-6 Man b1-3 Gal b1-
Glc a1-2 Gal a1-3 Man b1-
Gal a1-6 (Glc b1-2) Man a1-
Gal a1-2 (Man b1-6) Glc a1-
Glc b1-4 (Man a1-3) Gal a1-
Gal a1-3 Glc a1-2 Man a1-
Gal a1-4 Glc a1-6 Man b1-
Gal a1-6 Man b1-4 Glc a1-
Man b1-2 Glc a1-3 Gal a1-
Man b1-3 Glc b1-2 Gal b1-
Man b1-4 Gal b1-6 Glc a1-
Glc b1-6 Man a1-4 Gal a1-
Glc a1-2 Man b1-4 Gal b1-
Glc a1-3 Gal a1-3 Man b1-
Gal a1-6 (Glc b1-3) Man a1-
Gal a1-3 (Man b1-2) Glc a1-
Glc b1-4 (Man a1-6) Gal a1-
Gal a1-4 Glc a1-2 Man a1-
Gal a1-6 Glc a1-6 Man b1-
Gal b1-2 Man b1-4 Glc a1-
Man b1-3 Glc a1-3 Gal a1-
Man b1-4 Glc b1-2 Gal b1-
Man b1-6 Gal b1-6 Glc a1-
Glc a1-2 Man a1-6 Gal a1-
Glc a1-3 Man b1-4 Gal b1-
Glc a1-4 Gal a1-3 Man b1-
Gal a1-6 (Glc b1-4) Man a1-
Gal a1-3 (Man b1-4) Glc a1-
Glc b1-6 (Man a1-2) Gal a1-
Gal a1-6 Glc a1-2 Man a1-
Gal b1-2 Glc a1-6 Man b1-
Gal b1-3 Man b1-4 Glc a1-
Man b1-4 Glc a1-3 Gal a1-
Man b1-6 Glc b1-2 Gal b1-
Man a1-2 Gal a1-2 Glc b1
Glc a1-3 Man a1-6 Gal a1-
Glc a1-4 Man b1-4 Gal b1-
Glc a1-6 Gal a1-3 Man b1-
Gal b1-2 (Glc b1-3) Man a1-
Gal a1-3 (Man b1-6) Glc a1-
Glc b1-6 (Man a1-3) Gal a1-
Gal b1-2 Glc a1-2 Man a1-
Gal b1-3 Glc a1-6 Man b1-
Gal b1-4 Man b1-4 Glc a1-
Man b1-6 Glc a1-3 Gal a1-
Man a1-2 Glc b1-3 Gal b1-
Man a1-3 Gal a1-2 Glc b1
Glc a1-4 Man a1-6 Gal a1-
Glc a1-6 Man b1-4 Gal b1-
Glc b1-2 Gal a1-3 Man b1-
Gal b1-2 (Glc b1-4) Man a1-
Gal a1-4 (Man b1-2) Glc a1-
Glc b1-6 (Man a1-4) Gal a1-
Gal b1-3 Glc a1-2 Man a1-
Gal b1-4 Glc a1-6 Man b1-
Gal b1-6 Man b1-4 Glc a1-
Man a1-2 Glc a1-4 Gal a1-
Man a1-3 Glc b1-3 Gal b1-
Man a1-4 Gal a1-2 Glc b1
Glc a1-6 Man a1-6 Gal a1-
Glc b1-2 Man b1-4 Gal b1-
Glc b1-3 Gal a1-3 Man b1-
Gal b1-2 (Glc b1-6) Man a1-
Gal a1-4 (Man b1-3) Glc a1-
Glc a1-2 (Man b1-3) Gal a1-
Gal b1-4 Glc a1-2 Man a1-
Gal b1-6 Glc a1-6 Man b1-
Gal a1-2 Man b1-6 Glc a1-
Man a1-3 Glc a1-4 Gal a1-
Man a1-4 Glc b1-3 Gal b1-
Man a1-6 Gal a1-2 Glc b1
Glc b1-2 Man a1-6 Gal a1-
Glc b1-3 Man b1-4 Gal b1-
Glc b1-4 Gal a1-3 Man b1-
Gal b1-3 (Glc b1-2) Man a1-
Gal a1-4 (Man b1-6) Glc a1-
Glc a1-2 (Man b1-4) Gal a1-
Gal b1-6 Glc a1-2 Man a1-
Gal a1-2 Glc b1-2 Man b1-
Gal a1-3 Man b1-6 Glc a1-
Man a1-4 Glc a1-4 Gal a1-
Man a1-6 Glc b1-3 Gal b1-
Man b1-2 Gal a1-2 Glc b1
Glc b1-3 Man a1-6 Gal a1-
Glc b1-4 Man b1-4 Gal b1-
Glc b1-6 Gal a1-3 Man b1-
Gal b1-3 (Glc b1-4) Man a1-
Gal a1-6 (Man b1-2) Glc a1-
Glc a1-2 (Man b1-6) Gal a1-
Gal a1-2 Glc a1-3 Man a1-
Gal a1-3 Glc b1-2 Man b1-
Gal a1-4 Man b1-6 Glc a1-
Man a1-6 Glc a1-4 Gal a1-
Man b1-2 Glc b1-3 Gal b1-
Man b1-3 Gal a1-2 Glc b1
Glc b1-4 Man a1-6 Gal a1-
Glc b1-6 Man b1-4 Gal b1-
Glc a1-2 Gal a1-4 Man b1-
Gal b1-3 (Glc b1-6) Man a1-
Gal a1-6 (Man b1-3) Glc a1-
Glc a1-3 (Man b1-2) Gal a1-
Gal a1-3 Glc a1-3 Man a1-
Gal a1-4 Glc b1-2 Man b1-
Gal a1-6 Man b1-6 Glc a1-
Man b1-2 Glc a1-4 Gal a1-
Man b1-3 Glc b1-3 Gal b1-
Man b1-4 Gal a1-2 Glc b1
Glc b1-6 Man a1-6 Gal a1-
Glc a1-2 Man b1-6 Gal b1-
Glc a1-3 Gal a1-4 Man b1-
Gal b1-4 (Glc b1-2) Man a1-
Gal a1-6 (Man b1-4) Glc a1-
Glc a1-3 (Man b1-4) Gal a1-
Gal a1-4 Glc a1-3 Man a1-
Gal a1-6 Glc b1-2 Man b1-
Gal b1-2 Man b1-6 Glc a1-
Man b1-3 Glc a1-4 Gal a1-
Man b1-4 Glc b1-3 Gal b1-
Man b1-6 Gal a1-2 Glc b1
Glc a1-2 Man b1-2 Gal a1-
Glc a1-3 Man b1-6 Gal b1-
Glc a1-4 Gal a1-4 Man b1-
Gal b1-4 (Glc b1-3) Man a1-
Gal b1-2 (Man b1-3) Glc a1-
Glc a1-3 (Man b1-6) Gal a1-
Gal a1-6 Glc a1-3 Man a1-
Gal b1-2 Glc b1-2 Man b1-
Gal b1-3 Man b1-6 Glc a1-
Man b1-4 Glc a1-4 Gal a1-
Man b1-6 Glc b1-3 Gal b1-
Man a1-2 Gal a1-3 Glc b1
Glc a1-3 Man b1-2 Gal a1-
Glc a1-4 Man b1-6 Gal b1-
Glc a1-6 Gal a1-4 Man b1-
Gal b1-4 (Glc b1-6) Man a1-
Gal b1-2 (Man b1-4) Glc a1-
Glc a1-4 (Man b1-2) Gal a1-
Gal b1-2 Glc a1-3 Man a1-
Gal b1-3 Glc b1-2 Man b1-
Gal b1-4 Man b1-6 Glc a1-
Man b1-6 Glc a1-4 Gal a1-
Man a1-2 Glc b1-4 Gal b1-
Man a1-3 Gal a1-3 Glc b1
Glc a1-4 Man b1-2 Gal a1-
Glc a1-6 Man b1-6 Gal b1-
Glc b1-2 Gal a1-4 Man b1-
Gal b1-6 (Glc b1-2) Man a1-
Gal b1-2 (Man b1-6) Glc a1-
Glc a1-4 (Man b1-3) Gal a1-
Gal b1-3 Glc a1-3 Man a1-
Gal b1-4 Glc b1-2 Man b1-
Gal b1-6 Man b1-6 Glc a1-
Man a1-2 Glc a1-6 Gal a1-
Man a1-3 Glc b1-4 Gal b1-
Man a1-4 Gal a1-3 Glc b1
Glc a1-6 Man b1-2 Gal a1-
Glc b1-2 Man b1-6 Gal b1-
Glc b1-3 Gal a1-4 Man b1-
Gal b1-6 (Glc b1-3) Man a1-
Gal b1-3 (Man b1-2) Glc a1-
Glc a1-4 (Man b1-6) Gal a1-
Gal b1-4 Glc a1-3 Man a1-
Gal b1-6 Glc b1-2 Man b1-
Gal a1-2 Man a1-2 Glc b1-
Man a1-3 Glc a1-6 Gal a1-
Man a1-4 Glc b1-4 Gal b1-
Man a1-6 Gal a1-3 Glc b1
Glc b1-2 Man b1-2 Gal a1-
Glc b1-3 Man b1-6 Gal b1-
Glc b1-4 Gal a1-4 Man b1-
Gal b1-6 (Glc b1-4) Man a1-
Gal b1-3 (Man b1-4) Glc a1-
Glc a1-6 (Man b1-2) Gal a1-
Gal b1-6 Glc a1-3 Man a1-
Gal a1-2 Glc b1-3 Man b1-
Gal a1-3 Man a1-2 Glc b1-
Man a1-4 Glc a1-6 Gal a1-
Man a1-6 Glc b1-4 Gal b1-
Man b1-2 Gal a1-3 Glc b1
Glc b1-3 Man b1-2 Gal a1-
Glc b1-4 Man b1-6 Gal b1-
Glc b1-6 Gal a1-4 Man b1-
Gal a1-2 (Glc a1-3) Man b1-
Gal b1-3 (Man b1-6) Glc a1-
Glc a1-6 (Man b1-3) Gal a1-
Gal a1-2 Glc a1-4 Man a1-
Gal a1-3 Glc b1-3 Man b1-
Gal a1-4 Man a1-2 Glc b1-
Man a1-6 Glc a1-6 Gal a1-
Man b1-2 Glc b1-4 Gal b1-
Man b1-3 Gal a1-3 Glc b1
Glc b1-4 Man b1-2 Gal a1-
Glc b1-6 Man b1-6 Gal b1-
Glc a1-2 Gal a1-6 Man b1-
Gal a1-2 (Glc a1-4) Man b1-
Gal b1-4 (Man b1-2) Glc a1-
Glc a1-6 (Man b1-4) Gal a1-
Gal a1-3 Glc a1-4 Man a1-
Gal a1-4 Glc b1-3 Man b1-
Gal a1-6 Man a1-2 Glc b1-
Man b1-2 Glc a1-6 Gal a1-
Man b1-3 Glc b1-4 Gal b1-
Man b1-4 Gal a1-3 Glc b1
Glc b1-6 Man b1-2 Gal a1-
Glc a1-2 Gal a1-2 Man a1-
Glc a1-3 Gal a1-6 Man b1-
Gal a1-2 (Glc a1-6) Man b1-
Gal b1-4 (Man b1-3) Glc a1-
Glc b1-2 (Man b1-3) Gal a1-
Gal a1-4 Glc a1-4 Man a1-
Gal a1-6 Glc b1-3 Man b1-
Gal b1-2 Man a1-2 Glc b1-
Man b1-3 Glc a1-6 Gal a1-
Man b1-4 Glc b1-4 Gal b1-
Man b1-6 Gal a1-3 Glc b1
Glc a1-2 Man b1-3 Gal a1-
Glc a1-3 Gal a1-2 Man a1-
Glc a1-4 Gal a1-6 Man b1-
Gal a1-3 (Glc a1-2) Man b1-
Gal b1-4 (Man b1-6) Glc a1-
Glc b1-2 (Man b1-4) Gal a1-
Gal a1-6 Glc a1-4 Man a1-
Gal b1-2 Glc b1-3 Man b1-
Gal b1-3 Man a1-2 Glc b1-
Man b1-4 Glc a1-6 Gal a1-
Man b1-6 Glc b1-4 Gal b1-
Man a1-2 Gal a1-4 Glc b1
Glc a1-3 Man b1-3 Gal a1-
Glc a1-4 Gal a1-2 Man a1-
Glc a1-6 Gal a1-6 Man b1-
Gal a1-3 (Glc a1-4) Man b1-
Gal b1-6 (Man b1-2) Glc a1-
Glc b1-2 (Man b1-6) Gal a1-
Gal b1-2 Glc a1-4 Man a1-
Gal b1-3 Glc b1-3 Man b1-
Gal b1-4 Man a1-2 Glc b1-
Man b1-6 Glc a1-6 Gal a1-
Man a1-2 Glc b1-6 Gal b1-
Man a1-3 Gal a1-4 Glc b1
Glc a1-4 Man b1-3 Gal a1-
Glc a1-6 Gal a1-2 Man a1-
Glc b1-2 Gal a1-6 Man b1-
Gal a1-3 (Glc a1-6) Man b1-
Gal b1-6 (Man b1-3) Glc a1-
Glc b1-3 (Man b1-2) Gal a1-
Gal b1-3 Glc a1-4 Man a1-
Gal b1-4 Glc b1-3 Man b1-
Gal b1-6 Man a1-2 Glc b1-
Man a1-2 Glc b1-2 Gal a1-
Man a1-3 Glc b1-6 Gal b1-
Man a1-4 Gal a1-4 Glc b1
Glc a1-6 Man b1-3 Gal a1-
Glc b1-2 Gal a1-2 Man a1-
Glc b1-3 Gal a1-6 Man b1-
Gal a1-4 (Glc a1-2) Man b1-
Gal b1-6 (Man b1-4) Glc a1-
Glc b1-3 (Man b1-4) Gal a1-
Gal b1-4 Glc a1-4 Man a1-
Gal b1-6 Glc b1-3 Man b1-
Gal a1-2 Man a1-3 Glc b1-
Man a1-3 Glc b1-2 Gal a1-
Man a1-4 Glc b1-6 Gal b1-
Man a1-6 Gal a1-4 Glc b1
Glc b1-2 Man b1-3 Gal a1-
Glc b1-3 Gal a1-2 Man a1-
Glc b1-4 Gal a1-6 Man b1-
Gal a1-4 (Glc a1-3) Man b1-
Gal a1-2 (Man a1-3) Glc b1-
Glc b1-3 (Man b1-6) Gal a1-
Gal b1-6 Glc a1-4 Man a1-
Gal a1-2 Glc b1-4 Man b1-
Gal a1-3 Man a1-3 Glc b1-
Man a1-4 Glc b1-2 Gal a1-
Man a1-6 Glc b1-6 Gal b1-
Man b1-2 Gal a1-4 Glc b1
Glc b1-3 Man b1-3 Gal a1-
Glc b1-4 Gal a1-2 Man a1-
Glc b1-6 Gal a1-6 Man b1-
Gal a1-4 (Glc a1-6) Man b1-
Gal a1-2 (Man a1-4) Glc b1-
Glc b1-4 (Man b1-2) Gal a1-
Gal a1-2 Glc a1-6 Man a1-
Gal a1-3 Glc b1-4 Man b1-
Gal a1-4 Man a1-3 Glc b1-
Man a1-6 Glc b1-2 Gal a1-
Man b1-2 Glc b1-6 Gal b1-
Man b1-3 Gal a1-4 Glc b1
Glc b1-4 Man b1-3 Gal a1-
Glc b1-6 Gal a1-2 Man a1-
Glc a1-2 Gal b1-2 Man b1-
Gal a1-6 (Glc a1-2) Man b1-
Gal a1-2 (Man a1-6) Glc b1-
Glc b1-4 (Man b1-3) Gal a1-
Gal a1-3 Glc a1-6 Man a1-
Gal a1-4 Glc b1-4 Man b1-
Gal a1-6 Man a1-3 Glc b1-
Man b1-2 Glc b1-2 Gal a1-
Man b1-3 Glc b1-6 Gal b1-
Man b1-4 Gal a1-4 Glc b1
Glc b1-6 Man b1-3 Gal a1-
Glc a1-2 Gal a1-3 Man a1-
Glc a1-3 Gal b1-2 Man b1-
Gal a1-6 (Glc a1-3) Man b1-
Gal a1-3 (Man a1-2) Glc b1-
Glc b1-4 (Man b1-6) Gal a1-
Gal a1-4 Glc a1-6 Man a1-
Gal a1-6 Glc b1-4 Man b1-
Gal b1-2 Man a1-3 Glc b1-
Man b1-3 Glc b1-2 Gal a1-
Man b1-4 Glc b1-6 Gal b1-
Man b1-6 Gal a1-4 Glc b1
Glc a1-2 Man b1-4 Gal a1-
Glc a1-3 Gal a1-3 Man a1-
Glc a1-4 Gal b1-2 Man b1-
Gal a1-6 (Glc a1-4) Man b1-
Gal a1-3 (Man a1-4) Glc b1-
Glc b1-6 (Man b1-2) Gal a1-
Gal a1-6 Glc a1-6 Man a1-
Gal b1-2 Glc b1-4 Man b1-
Gal b1-3 Man a1-3 Glc b1-
Man b1-4 Glc b1-2 Gal a1-
Man b1-6 Glc b1-6 Gal b1-
Man a1-2 Gal a1-6 Glc b1
Glc a1-3 Man b1-4 Gal a1-
Glc a1-4 Gal a1-3 Man a1-
Glc a1-6 Gal b1-2 Man b1-
Gal b1-2 (Glc a1-3) Man b1-
Gal a1-3 (Man a1-6) Glc b1-
Glc b1-6 (Man b1-3) Gal a1-
Gal b1-2 Glc a1-6 Man a1-
Gal b1-3 Glc b1-4 Man b1-
Gal b1-4 Man a1-3 Glc b1-
Man b1-6 Glc b1-2 Gal a1-
Man a1-2 Gal a1-2 Glc a1-
Man a1-3 Gal a1-6 Glc b1
Glc a1-4 Man b1-4 Gal a1-
Glc a1-6 Gal a1-3 Man a1-
Glc b1-2 Gal b1-2 Man b1-
Gal b1-2 (Glc a1-4) Man b1-
Gal a1-4 (Man a1-2) Glc b1-
Glc b1-6 (Man b1-4) Gal a1-
Gal b1-3 Glc a1-6 Man a1-
Gal b1-4 Glc b1-4 Man b1-
Gal b1-6 Man a1-3 Glc b1-
Man a1-2 Glc b1-3 Gal a1-
Man a1-3 Gal a1-2 Glc a1-
Man a1-4 Gal a1-6 Glc b1
Glc a1-6 Man b1-4 Gal a1-
Glc b1-2 Gal a1-3 Man a1-
Glc b1-3 Gal b1-2 Man b1-
Gal b1-2 (Glc a1-6) Man b1-
Gal a1-4 (Man a1-3) Glc b1-
Glc a1-2 (Man a1-3) Gal b1-
Gal b1-4 Glc a1-6 Man a1-
Gal b1-6 Glc b1-4 Man b1-
Gal a1-2 Man a1-4 Glc b1-
Man a1-3 Glc b1-3 Gal a1-
Man a1-4 Gal a1-2 Glc a1-
Man a1-6 Gal a1-6 Glc b1
Glc b1-2 Man b1-4 Gal a1-
Glc b1-3 Gal a1-3 Man a1-
Glc b1-4 Gal b1-2 Man b1-
Gal b1-3 (Glc a1-2) Man b1-
Gal a1-4 (Man a1-6) Glc b1-
Glc a1-2 (Man a1-4) Gal b1-
Gal b1-6 Glc a1-6 Man a1-
Gal a1-2 Glc b1-6 Man b1-
Gal a1-3 Man a1-4 Glc b1-
Man a1-4 Glc b1-3 Gal a1-
Man a1-6 Gal a1-2 Glc a1-
Man b1-2 Gal a1-6 Glc b1
Glc b1-3 Man b1-4 Gal a1-
Glc b1-4 Gal a1-3 Man a1-
Glc b1-6 Gal b1-2 Man b1-
Gal b1-3 (Glc a1-4) Man b1-
Gal a1-6 (Man a1-2) Glc b1-
Glc a1-2 (Man a1-6) Gal b1-
Gal a1-2 Glc b1-2 Man a1-
Gal a1-3 Glc b1-6 Man b1-
Gal a1-4 Man a1-4 Glc b1-
Man a1-6 Glc b1-3 Gal a1-
Man b1-2 Gal a1-2 Glc a1-
Man b1-3 Gal a1-6 Glc b1
Glc b1-4 Man b1-4 Gal a1-
Glc b1-6 Gal a1-3 Man a1-
Glc a1-2 Gal b1-3 Man b1-
Gal b1-3 (Glc a1-6) Man b1-
Gal a1-6 (Man a1-3) Glc b1-
Glc a1-3 (Man a1-2) Gal b1-
Gal a1-3 Glc b1-2 Man a1-
Gal a1-4 Glc b1-6 Man b1-
Gal a1-6 Man a1-4 Glc b1-
Man b1-2 Glc b1-3 Gal a1-
Man b1-3 Gal a1-2 Glc a1-
Man b1-4 Gal a1-6 Glc b1
Glc b1-6 Man b1-4 Gal a1-
Glc a1-2 Gal a1-4 Man a1-
Glc a1-3 Gal b1-3 Man b1-
Gal b1-4 (Glc a1-2) Man b1-
Gal a1-6 (Man a1-4) Glc b1-
Glc a1-3 (Man a1-4) Gal b1-
Gal a1-4 Glc b1-2 Man a1-
Gal a1-6 Glc b1-6 Man b1-
Gal b1-2 Man a1-4 Glc b1-
Man b1-3 Glc b1-3 Gal a1-
Man b1-4 Gal a1-2 Glc a1-
Man b1-6 Gal a1-6 Glc b1
Glc a1-2 Man b1-6 Gal a1-
Glc a1-3 Gal a1-4 Man a1-
Glc a1-4 Gal b1-3 Man b1-
Gal b1-4 (Glc a1-3) Man b1-
Gal b1-2 (Man a1-3) Glc b1-
Glc a1-3 (Man a1-6) Gal b1-
Gal a1-6 Glc b1-2 Man a1-
Gal b1-2 Glc b1-6 Man b1-
Gal b1-3 Man a1-4 Glc b1-
Man b1-4 Glc b1-3 Gal a1-
Man b1-6 Gal a1-2 Glc a1-
Man a1-2 Gal b1-2 Glc b1
Glc a1-3 Man b1-6 Gal a1-
Glc a1-4 Gal a1-4 Man a1-
Glc a1-6 Gal b1-3 Man b1-
Gal b1-4 (Glc a1-6) Man b1-
Gal b1-2 (Man a1-4) Glc b1-
Glc a1-4 (Man a1-2) Gal b1-
Gal b1-2 Glc b1-2 Man a1-
Gal b1-3 Glc b1-6 Man b1-
Gal b1-4 Man a1-4 Glc b1-
Man b1-6 Glc b1-3 Gal a1-
Man a1-2 Gal a1-3 Glc a1-
Man a1-3 Gal b1-2 Glc b1
Glc a1-4 Man b1-6 Gal a1-
Glc a1-6 Gal a1-4 Man a1-
Glc b1-2 Gal b1-3 Man b1-
Gal b1-6 (Glc a1-2) Man b1-
Gal b1-2 (Man a1-6) Glc b1-
Glc a1-4 (Man a1-3) Gal b1-
Gal b1-3 Glc b1-2 Man a1-
Gal b1-4 Glc b1-6 Man b1-
Gal b1-6 Man a1-4 Glc b1-
Man a1-2 Glc b1-4 Gal a1-
Man a1-3 Gal a1-3 Glc a1-
Man a1-4 Gal b1-2 Glc b1
Glc a1-6 Man b1-6 Gal a1-
Glc b1-2 Gal a1-4 Man a1-
Glc b1-3 Gal b1-3 Man b1-
Gal b1-6 (Glc a1-3) Man b1-
Gal b1-3 (Man a1-2) Glc b1-
Glc a1-4 (Man a1-6) Gal b1-
Gal b1-4 Glc b1-2 Man a1-
Gal b1-6 Glc b1-6 Man b1-
Gal a1-2 Man a1-6 Glc b1-
Man a1-3 Glc b1-4 Gal a1-
Man a1-4 Gal a1-3 Glc a1-
Man a1-6 Gal b1-2 Glc b1
Glc b1-2 Man b1-6 Gal a1-
Glc b1-3 Gal a1-4 Man a1-
Glc b1-4 Gal b1-3 Man b1-
Gal b1-6 (Glc a1-4) Man b1-
Gal b1-3 (Man a1-4) Glc b1-
Glc a1-6 (Man a1-2) Gal b1-
Gal b1-6 Glc b1-2 Man a1-
Gal a1-2 Man a1-2 Glc a1-
Gal a1-3 Man a1-6 Glc b1-
Man a1-4 Glc b1-4 Gal a1-
Man a1-6 Gal a1-3 Glc a1-
Man b1-2 Gal b1-2 Glc b1
Glc b1-3 Man b1-6 Gal a1-
Glc b1-4 Gal a1-4 Man a1-
Glc b1-6 Gal b1-3 Man b1-
Gal a1-2 (Glc b1-3) Man b1-
Gal b1-3 (Man a1-6) Glc b1-
Glc a1-6 (Man a1-3) Gal b1-
Gal a1-2 Glc b1-3 Man a1-
Gal a1-3 Man a1-2 Glc a1-
Gal a1-4 Man a1-6 Glc b1-
Man a1-6 Glc b1-4 Gal a1-
Man b1-2 Gal a1-3 Glc a1-
Man b1-3 Gal b1-2 Glc b1
Glc b1-4 Man b1-6 Gal a1-
Glc b1-6 Gal a1-4 Man a1-
Glc a1-2 Gal b1-4 Man b1-
Gal a1-2 (Glc b1-4) Man b1-
Gal b1-4 (Man a1-2) Glc b1-
Glc a1-6 (Man a1-4) Gal b1-
Gal a1-3 Glc b1-3 Man a1-
Gal a1-4 Man a1-2 Glc a1-
Gal a1-6 Man a1-6 Glc b1-
Man b1-2 Glc b1-4 Gal a1-
Man b1-3 Gal a1-3 Glc a1-
Man b1-4 Gal b1-2 Glc b1
Glc b1-6 Man b1-6 Gal a1-
Glc a1-2 Gal a1-6 Man a1-
Glc a1-3 Gal b1-4 Man b1-
Gal a1-2 (Glc b1-6) Man b1-
Gal b1-4 (Man a1-3) Glc b1-
Glc b1-2 (Man a1-3) Gal b1-
Gal a1-4 Glc b1-3 Man a1-
Gal a1-6 Man a1-2 Glc a1-
Gal b1-2 Man a1-6 Glc b1-
Man b1-3 Glc b1-4 Gal a1-
Man b1-4 Gal a1-3 Glc a1-
Man b1-6 Gal b1-2 Glc b1
Glc a1-2 Man a1-2 Gal b1-
Glc a1-3 Gal a1-6 Man a1-
Glc a1-4 Gal b1-4 Man b1-
Gal a1-3 (Glc b1-2) Man b1-
Gal b1-4 (Man a1-6) Glc b1-
Glc b1-2 (Man a1-4) Gal b1-
Gal a1-6 Glc b1-3 Man a1-
Gal b1-2 Man a1-2 Glc a1-
Gal b1-3 Man a1-6 Glc b1-
Man b1-4 Glc b1-4 Gal a1-
Man b1-6 Gal a1-3 Glc a1-
Man a1-2 Gal b1-3 Glc b1
Glc a1-3 Man a1-2 Gal b1-
Glc a1-4 Gal a1-6 Man a1-
Glc a1-6 Gal b1-4 Man b1-
Gal a1-3 (Glc b1-4) Man b1-
Gal b1-6 (Man a1-2) Glc b1-
Glc b1-2 (Man a1-6) Gal b1-
Gal b1-2 Glc b1-3 Man a1-
Gal b1-3 Man a1-2 Glc a1-
Gal b1-4 Man a1-6 Glc b1-
Man b1-6 Glc b1-4 Gal a1-
Man a1-2 Gal a1-4 Glc a1-
Man a1-3 Gal b1-3 Glc b1
Glc a1-4 Man a1-2 Gal b1-
Glc a1-6 Gal a1-6 Man a1-
Glc b1-2 Gal b1-4 Man b1-
Gal a1-3 (Glc b1-6) Man b1-
Gal b1-6 (Man a1-3) Glc b1-
Glc b1-3 (Man a1-2) Gal b1-
Gal b1-3 Glc b1-3 Man a1-
Gal b1-4 Man a1-2 Glc a1-
Gal b1-6 Man a1-6 Glc b1-
Man a1-2 Glc b1-6 Gal a1-
Man a1-3 Gal a1-4 Glc a1-
Man a1-4 Gal b1-3 Glc b1
Glc a1-6 Man a1-2 Gal b1-
Glc b1-2 Gal a1-6 Man a1-
Glc b1-3 Gal b1-4 Man b1-
Gal a1-4 (Glc b1-2) Man b1-
Gal b1-6 (Man a1-4) Glc b1-
Glc b1-3 (Man a1-4) Gal b1-
Gal b1-4 Glc b1-3 Man a1-
Gal b1-6 Man a1-2 Glc a1-
Gal a1-2 Man b1-2 Glc b1-
Man a1-3 Glc b1-6 Gal a1-
Man a1-4 Gal a1-4 Glc a1-
Man a1-6 Gal b1-3 Glc b1
Glc b1-2 Man a1-2 Gal b1-
Glc b1-3 Gal a1-6 Man a1-
Glc b1-4 Gal b1-4 Man b1-
Gal a1-4 (Glc b1-3) Man b1-
Gal a1-2 (Man b1-3) Glc b1-
Glc b1-3 (Man a1-6) Gal b1-
Gal b1-6 Glc b1-3 Man a1-
Gal a1-2 Man a1-3 Glc a1-
Gal a1-3 Man b1-2 Glc b1-
Man a1-4 Glc b1-6 Gal a1-
Man a1-6 Gal a1-4 Glc a1-
Man b1-2 Gal b1-3 Glc b1
Glc b1-3 Man a1-2 Gal b1-
Glc b1-4 Gal a1-6 Man a1-
Glc b1-6 Gal b1-4 Man b1-
Gal a1-4 (Glc b1-6) Man b1-
Gal a1-2 (Man b1-4) Glc b1-
Glc b1-4 (Man a1-2) Gal b1-
Gal a1-2 Glc b1-4 Man a1-
Gal a1-3 Man a1-3 Glc a1-
Gal a1-4 Man b1-2 Glc b1-
Man a1-6 Glc b1-6 Gal a1-
Man b1-2 Gal a1-4 Glc a1-
Man b1-3 Gal b1-3 Glc b1
Glc b1-4 Man a1-2 Gal b1-
Glc b1-6 Gal a1-6 Man a1-
Glc a1-2 Gal b1-6 Man b1-
Gal a1-6 (Glc b1-2) Man b1-
Gal a1-2 (Man b1-6) Glc b1-
Glc b1-4 (Man a1-3) Gal b1-
Gal a1-3 Glc b1-4 Man a1-
Gal a1-4 Man a1-3 Glc a1-
Gal a1-6 Man b1-2 Glc b1-
Man b1-2 Glc b1-6 Gal a1-
Man b1-3 Gal a1-4 Glc a1-
Man b1-4 Gal b1-3 Glc b1
Glc b1-6 Man a1-2 Gal b1-
Glc a1-2 Gal b1-2 Man a1-
Glc a1-3 Gal b1-6 Man b1-
Gal a1-6 (Glc b1-3) Man b1-
Gal a1-3 (Man b1-2) Glc b1-
Glc b1-4 (Man a1-6) Gal b1-
Gal a1-4 Glc b1-4 Man a1-
Gal a1-6 Man a1-3 Glc a1-
Gal b1-2 Man b1-2 Glc b1-
Man b1-3 Glc b1-6 Gal a1-
Man b1-4 Gal a1-4 Glc a1-
Man b1-6 Gal b1-3 Glc b1
Glc a1-2 Man a1-3 Gal b1-
Glc a1-3 Gal b1-2 Man a1-
Glc a1-4 Gal b1-6 Man b1-
Gal a1-6 (Glc b1-4) Man b1-
Gal a1-3 (Man b1-4) Glc b1-
Glc b1-6 (Man a1-2) Gal b1-
Gal a1-6 Glc b1-4 Man a1-
Gal b1-2 Man a1-3 Glc a1-
Gal b1-3 Man b1-2 Glc b1-
Man b1-4 Glc b1-6 Gal a1-
Man b1-6 Gal a1-4 Glc a1-
Man a1-2 Gal b1-4 Glc b1
Glc a1-3 Man a1-3 Gal b1-
Glc a1-4 Gal b1-2 Man a1-
Glc a1-6 Gal b1-6 Man b1-
Gal b1-2 (Glc b1-3) Man b1-
Gal a1-3 (Man b1-6) Glc b1-
Glc b1-6 (Man a1-3) Gal b1-
Gal b1-2 Glc b1-4 Man a1-
Gal b1-3 Man a1-3 Glc a1-
Gal b1-4 Man b1-2 Glc b1-
Man b1-6 Glc b1-6 Gal a1-
Man a1-2 Gal a1-6 Glc a1-
Man a1-3 Gal b1-4 Glc b1
Glc a1-4 Man a1-3 Gal b1-
Glc a1-6 Gal b1-2 Man a1-
Glc b1-2 Gal b1-6 Man b1-
Gal b1-2 (Glc b1-4) Man b1-
Gal a1-4 (Man b1-2) Glc b1-
Glc b1-6 (Man a1-4) Gal b1-
Gal b1-3 Glc b1-4 Man a1-
Gal b1-4 Man a1-3 Glc a1-
Gal b1-6 Man b1-2 Glc b1-
Man a1-2 Glc a1-2 Gal b1-
Man a1-3 Gal a1-6 Glc a1-
Man a1-4 Gal b1-4 Glc b1
Glc a1-6 Man a1-3 Gal b1-
Glc b1-2 Gal b1-2 Man a1-
Glc b1-3 Gal b1-6 Man b1-
Gal b1-2 (Glc b1-6) Man b1-
Gal a1-4 (Man b1-3) Glc b1-
Glc a1-2 (Man b1-3) Gal b1-
Gal b1-4 Glc b1-4 Man a1-
Gal b1-6 Man a1-3 Glc a1-
Gal a1-2 Man b1-3 Glc b1-
Man a1-3 Glc a1-2 Gal b1-
Man a1-4 Gal a1-6 Glc a1-
Man a1-6 Gal b1-4 Glc b1
Glc b1-2 Man a1-3 Gal b1-
Glc b1-3 Gal b1-2 Man a1-
Glc b1-4 Gal b1-6 Man b1-
Gal b1-3 (Glc b1-2) Man b1-
Gal a1-4 (Man b1-6) Glc b1-
Glc a1-2 (Man b1-4) Gal b1-
Gal b1-6 Glc b1-4 Man a1-
Gal a1-2 Man a1-4 Glc a1-
Gal a1-3 Man b1-3 Glc b1-
Man a1-4 Glc a1-2 Gal b1-
Man a1-6 Gal a1-6 Glc a1-
Man b1-2 Gal b1-4 Glc b1
Glc b1-3 Man a1-3 Gal b1-
Glc b1-4 Gal b1-2 Man a1-
Glc b1-6 Gal b1-6 Man b1-
Gal b1-3 (Glc b1-4) Man b1-
Gal a1-6 (Man b1-2) Glc b1-
Glc a1-2 (Man b1-6) Gal b1-
Gal a1-2 Glc b1-6 Man a1-
Gal a1-3 Man a1-4 Glc a1-
Gal a1-4 Man b1-3 Glc b1-
Man a1-6 Glc a1-2 Gal b1-
Man b1-2 Gal a1-6 Glc a1-
Man b1-3 Gal b1-4 Glc b1
Glc b1-4 Man a1-3 Gal b1-
Glc b1-6 Gal b1-2 Man a1-
Gal a1-2 (Glc a1-3) Man a1-
Gal b1-3 (Glc b1-6) Man b1-
Gal a1-6 (Man b1-3) Glc b1-
Glc a1-3 (Man b1-2) Gal b1-
Gal a1-3 Glc b1-6 Man a1-
Gal a1-4 Man a1-4 Glc a1-
Gal a1-6 Man b1-3 Glc b1-
Man b1-2 Glc a1-2 Gal b1-
Man b1-3 Gal a1-6 Glc a1-
Man b1-4 Gal b1-4 Glc b1
Glc b1-6 Man a1-3 Gal b1-
Glc a1-2 Gal b1-3 Man a1-
Gal a1-2 (Glc a1-4) Man a1-
Gal b1-4 (Glc b1-2) Man b1-
Gal a1-6 (Man b1-4) Glc b1-
Glc a1-3 (Man b1-4) Gal b1-
Gal a1-4 Glc b1-6 Man a1-
Gal a1-6 Man a1-4 Glc a1-
Gal b1-2 Man b1-3 Glc b1-
Man b1-3 Glc a1-2 Gal b1-
Man b1-4 Gal a1-6 Glc a1-
Man b1-6 Gal b1-4 Glc b1
Glc a1-2 Man a1-4 Gal b1-
Glc a1-3 Gal b1-3 Man a1-
Gal a1-2 (Glc a1-6) Man a1-
Gal b1-4 (Glc b1-3) Man b1-
Gal b1-2 (Man b1-3) Glc b1-
Glc a1-3 (Man b1-6) Gal b1-
Gal a1-6 Glc b1-6 Man a1-
Gal b1-2 Man a1-4 Glc a1-
Gal b1-3 Man b1-3 Glc b1-
Man b1-4 Glc a1-2 Gal b1-
Man b1-6 Gal a1-6 Glc a1-
Man a1-2 Gal b1-6 Glc b1
Glc a1-3 Man a1-4 Gal b1-
Glc a1-4 Gal b1-3 Man a1-
Gal a1-3 (Glc a1-2) Man a1-
Gal b1-4 (Glc b1-6) Man b1-
Gal b1-2 (Man b1-4) Glc b1-
Glc a1-4 (Man b1-2) Gal b1-
Gal b1-2 Glc b1-6 Man a1-
Gal b1-3 Man a1-4 Glc a1-
Gal b1-4 Man b1-3 Glc b1-
Man b1-6 Glc a1-2 Gal b1-
Man a1-2 Gal b1-2 Glc a1-
Man a1-3 Gal b1-6 Glc b1
Glc a1-4 Man a1-4 Gal b1-
Glc a1-6 Gal b1-3 Man a1-
Gal a1-3 (Glc a1-4) Man a1-
Gal b1-6 (Glc b1-2) Man b1-
Gal b1-2 (Man b1-6) Glc b1-
Glc a1-4 (Man b1-3) Gal b1-
Gal b1-3 Glc b1-6 Man a1-
Gal b1-4 Man a1-4 Glc a1-
Gal b1-6 Man b1-3 Glc b1-
Man a1-2 Glc a1-3 Gal b1-
Man a1-3 Gal b1-2 Glc a1-
Man a1-4 Gal b1-6 Glc b1
Glc a1-6 Man a1-4 Gal b1-
Glc b1-2 Gal b1-3 Man a1-
Gal a1-3 (Glc a1-6) Man a1-
Gal b1-6 (Glc b1-3) Man b1-
Gal b1-3 (Man b1-2) Glc b1-
Glc a1-4 (Man b1-6) Gal b1-
Gal b1-4 Glc b1-6 Man a1-
Gal b1-6 Man a1-4 Glc a1-
Gal a1-2 Man b1-4 Glc b1-
Man a1-3 Glc a1-3 Gal b1-
Man a1-4 Gal b1-2 Glc a1-
Man a1-6 Gal b1-6 Glc b1
Glc b1-2 Man a1-4 Gal b1-
Glc b1-3 Gal b1-3 Man a1-
Gal a1-4 (Glc a1-2) Man a1-
Gal b1-6 (Glc b1-4) Man b1-
Gal b1-3 (Man b1-4) Glc b1-
Glc a1-6 (Man b1-2) Gal b1-
Gal b1-6 Glc b1-6 Man a1-
Gal a1-2 Man a1-6 Glc a1-
Gal a1-3 Man b1-4 Glc b1-
Man a1-4 Glc a1-3 Gal b1-
Man a1-6 Gal b1-2 Glc a1-
Man b1-2 Gal b1-6 Glc b1
Glc b1-3 Man a1-4 Gal b1-
Glc b1-4 Gal b1-3 Man a1-
Gal a1-4 (Glc a1-3) Man a1-
Gal a1-2 (Man a1-3) Glc a1-
Gal b1-3 (Man b1-6) Glc b1-
Glc a1-6 (Man b1-3) Gal b1-
Gal a1-2 Glc a1-2 Man b1-
Gal a1-3 Man a1-6 Glc a1-
Gal a1-4 Man b1-4 Glc b1-
Man a1-6 Glc a1-3 Gal b1-
Man b1-2 Gal b1-2 Glc a1-
Man b1-3 Gal b1-6 Glc b1
Glc b1-4 Man a1-4 Gal b1-
Glc b1-6 Gal b1-3 Man a1-
Gal a1-4 (Glc a1-6) Man a1-
Gal a1-2 (Man a1-4) Glc a1-
Gal b1-4 (Man b1-2) Glc b1-
Glc a1-6 (Man b1-4) Gal b1-
Gal a1-3 Glc a1-2 Man b1-
Gal a1-4 Man a1-6 Glc a1-
Gal a1-6 Man b1-4 Glc b1-
Man b1-2 Glc a1-3 Gal b1-
Man b1-3 Gal b1-2 Glc a1-
Man b1-4 Gal b1-6 Glc b1
Glc b1-6 Man a1-4 Gal b1-
Glc a1-2 Gal b1-4 Man a1-
Gal a1-6 (Glc a1-2) Man a1-
Gal a1-2 (Man a1-6) Glc a1-
Gal b1-4 (Man b1-3) Glc b1-
Glc b1-2 (Man b1-3) Gal b1-
Gal a1-4 Glc a1-2 Man b1-
Gal a1-6 Man a1-6 Glc a1-
Gal b1-2 Man b1-4 Glc b1-
Man b1-3 Glc a1-3 Gal b1-
Man b1-4 Gal b1-2 Glc a1-
Man b1-6 Gal b1-6 Glc b1
Glc a1-2 Man a1-6 Gal b1-
Glc a1-3 Gal b1-4 Man a1-
Gal a1-6 (Glc a1-3) Man a1-
Gal a1-3 (Man a1-2) Glc a1-
Gal b1-4 (Man b1-6) Glc b1-
Glc b1-2 (Man b1-4) Gal b1-
Gal a1-6 Glc a1-2 Man b1-
Gal b1-2 Man a1-6 Glc a1-
Gal b1-3 Man b1-4 Glc b1-
Man b1-4 Glc a1-3 Gal b1-
Man b1-6 Gal b1-2 Glc a1-
Glc a1-2 Man a1-2 Gal a1-
Glc a1-3 Man a1-6 Gal b1-
Glc a1-4 Gal b1-4 Man a1-
Gal a1-6 (Glc a1-4) Man a1-
Gal a1-3 (Man a1-4) Glc a1-
Gal b1-6 (Man b1-2) Glc b1-
Glc b1-2 (Man b1-6) Gal b1-
Gal b1-2 Glc a1-2 Man b1-
Gal b1-3 Man a1-6 Glc a1-
Gal b1-4 Man b1-4 Glc b1-
Man b1-6 Glc a1-3 Gal b1-
Man a1-2 Gal b1-3 Glc a1-
Glc a1-3 Man a1-2 Gal a1-
Glc a1-4 Man a1-6 Gal b1-
Glc a1-6 Gal b1-4 Man a1-
Gal b1-2 (Glc a1-3) Man a1-
Gal a1-3 (Man a1-6) Glc a1-
Gal b1-6 (Man b1-3) Glc b1-
Glc b1-3 (Man b1-2) Gal b1-
Gal b1-3 Glc a1-2 Man b1-
Gal b1-4 Man a1-6 Glc a1-
Gal b1-6 Man b1-4 Glc b1-
Man a1-2 Glc a1-4 Gal b1-
Man a1-3 Gal b1-3 Glc a1-
Glc a1-4 Man a1-2 Gal a1-
Glc a1-6 Man a1-6 Gal b1-
Glc b1-2 Gal b1-4 Man a1-
Gal b1-2 (Glc a1-4) Man a1-
Gal a1-4 (Man a1-2) Glc a1-
Gal b1-6 (Man b1-4) Glc b1-
Glc b1-3 (Man b1-4) Gal b1-
Gal b1-4 Glc a1-2 Man b1-
Gal b1-6 Man a1-6 Glc a1-
Gal a1-2 Man b1-6 Glc b1-
Man a1-3 Glc a1-4 Gal b1-
Man a1-4 Gal b1-3 Glc a1-
Glc a1-6 Man a1-2 Gal a1-
Glc b1-2 Man a1-6 Gal b1-
Glc b1-3 Gal b1-4 Man a1-
Gal b1-2 (Glc a1-6) Man a1-
Gal a1-4 (Man a1-3) Glc a1-
Glc a1-2 (Man a1-3) Gal a1-
Glc b1-3 (Man b1-6) Gal b1-
Gal b1-6 Glc a1-2 Man b1-
Gal a1-2 Man b1-2 Glc a1-
Gal a1-3 Man b1-6 Glc b1-
Man a1-4 Glc a1-4 Gal b1-
Man a1-6 Gal b1-3 Glc a1-
Glc b1-2 Man a1-2 Gal a1-
Glc b1-3 Man a1-6 Gal b1-
Glc b1-4 Gal b1-4 Man a1-
Gal b1-3 (Glc a1-2) Man a1-
Gal a1-4 (Man a1-6) Glc a1-
Glc a1-2 (Man a1-4) Gal a1-
Glc b1-4 (Man b1-2) Gal b1-
Gal a1-2 Glc a1-3 Man b1-
Gal a1-3 Man b1-2 Glc a1-
Gal a1-4 Man b1-6 Glc b1-
Man a1-6 Glc a1-4 Gal b1-
Man b1-2 Gal b1-3 Glc a1-
Glc b1-3 Man a1-2 Gal a1-
Glc b1-4 Man a1-6 Gal b1-
Glc b1-6 Gal b1-4 Man a1-
Gal b1-3 (Glc a1-4) Man a1-
Gal a1-6 (Man a1-2) Glc a1-
Glc a1-2 (Man a1-6) Gal a1-
Glc b1-4 (Man b1-3) Gal b1-
Gal a1-3 Glc a1-3 Man b1-
Gal a1-4 Man b1-2 Glc a1-
Gal a1-6 Man b1-6 Glc b1-
Man b1-2 Glc a1-4 Gal b1-
Man b1-3 Gal b1-3 Glc a1-
Glc b1-4 Man a1-2 Gal a1-
Glc b1-6 Man a1-6 Gal b1-
Glc a1-2 Gal b1-6 Man a1-
Gal b1-3 (Glc a1-6) Man a1-
Gal a1-6 (Man a1-3) Glc a1-
Glc a1-3 (Man a1-2) Gal a1-
Glc b1-4 (Man b1-6) Gal b1-
Gal a1-4 Glc a1-3 Man b1-
Gal a1-6 Man b1-2 Glc a1-
Gal b1-2 Man b1-6 Glc b1-
Man b1-3 Glc a1-4 Gal b1-
Man b1-4 Gal b1-3 Glc a1-
Glc b1-6 Man a1-2 Gal a1-
Glc a1-2 Man b1-2 Gal b1-
Glc a1-3 Gal b1-6 Man a1-
Gal b1-4 (Glc a1-2) Man a1-
Gal a1-6 (Man a1-4) Glc a1-
Glc a1-3 (Man a1-4) Gal a1-
Glc b1-6 (Man b1-2) Gal b1-
Gal a1-6 Glc a1-3 Man b1-
Gal b1-2 Man b1-2 Glc a1-
Gal b1-3 Man b1-6 Glc b1-
Man b1-4 Glc a1-4 Gal b1-
Man b1-6 Gal b1-3 Glc a1-
Glc a1-2 Man a1-3 Gal a1-
Glc a1-3 Man b1-2 Gal b1-
Glc a1-4 Gal b1-6 Man a1-
Gal b1-4 (Glc a1-3) Man a1-
Gal b1-2 (Man a1-3) Glc a1-
Glc a1-3 (Man a1-6) Gal a1-
Glc b1-6 (Man b1-3) Gal b1-
Gal b1-2 Glc a1-3 Man b1-
Gal b1-3 Man b1-2 Glc a1-
Gal b1-4 Man b1-6 Glc b1-
Man b1-6 Glc a1-4 Gal b1-
Man a1-2 Gal b1-4 Glc a1-
Glc a1-3 Man a1-3 Gal a1-
Glc a1-4 Man b1-2 Gal b1-
Glc a1-6 Gal b1-6 Man a1-
Gal b1-4 (Glc a1-6) Man a1-
Gal b1-2 (Man a1-4) Glc a1-
Glc a1-4 (Man a1-2) Gal a1-
Glc b1-6 (Man b1-4) Gal b1-
Gal b1-3 Glc a1-3 Man b1-
Gal b1-4 Man b1-2 Glc a1-
Gal b1-6 Man b1-6 Glc b1-
Man a1-2 Glc a1-6 Gal b1-
Man a1-3 Gal b1-4 Glc a1-
Glc a1-4 Man a1-3 Gal a1-
Glc a1-6 Man b1-2 Gal b1-
Glc b1-2 Gal b1-6 Man a1-
Gal b1-6 (Glc a1-2) Man a1-
Gal b1-2 (Man a1-6) Glc a1-
Glc a1-4 (Man a1-3) Gal a1-
Gal b1-4 Glc a1-3 Man b1-
Gal b1-6 Man b1-2 Glc a1-
Man a1-2 Glc a1-2 Gal a1-
Man a1-3 Glc a1-6 Gal b1-
Man a1-4 Gal b1-4 Glc a1-
Glc a1-6 Man a1-3 Gal a1-
Glc b1-2 Man b1-2 Gal b1-
Glc b1-3 Gal b1-6 Man a1-
Gal b1-6 (Glc a1-3) Man a1-
Gal b1-3 (Man a1-2) Glc a1-
Glc a1-4 (Man a1-6) Gal a1-
Gal b1-6 Glc a1-3 Man b1-
Gal a1-2 Man b1-3 Glc a1-
Man a1-3 Glc a1-2 Gal a1-
Man a1-4 Glc a1-6 Gal b1-
Man a1-6 Gal b1-4 Glc a1-
Glc b1-2 Man a1-3 Gal a1-
Glc b1-3 Man b1-2 Gal b1-
Glc b1-4 Gal b1-6 Man a1-
Gal b1-6 (Glc a1-4) Man a1-
Gal b1-3 (Man a1-4) Glc a1-
Glc a1-6 (Man a1-2) Gal a1-
Gal a1-2 Glc a1-4 Man b1-
Gal a1-3 Man b1-3 Glc a1-
Man a1-4 Glc a1-2 Gal a1-
Man a1-6 Glc a1-6 Gal b1-
Man b1-2 Gal b1-4 Glc a1-
Glc b1-3 Man a1-3 Gal a1-
Glc b1-4 Man b1-2 Gal b1-
Glc b1-6 Gal b1-6 Man a1-
Gal a1-2 (Glc b1-3) Man a1-
Gal b1-3 (Man a1-6) Glc a1-
Glc a1-6 (Man a1-3) Gal a1-
Gal a1-3 Glc a1-4 Man b1-
Gal a1-4 Man b1-3 Glc a1-
Man a1-6 Glc a1-2 Gal a1-
Man b1-2 Glc a1-6 Gal b1-
Man b1-3 Gal b1-4 Glc a1-
Glc b1-4 Man a1-3 Gal a1-
Glc b1-6 Man b1-2 Gal b1-
Glc a1-2 Gal a1-2 Man b1-
Gal a1-2 (Glc b1-4) Man a1-
Gal b1-4 (Man a1-2) Glc a1-
Glc a1-6 (Man a1-4) Gal a1-
Gal a1-4 Glc a1-4 Man b1-
Gal a1-6 Man b1-3 Glc a1-
Man b1-2 Glc a1-2 Gal a1-
Man b1-3 Glc a1-6 Gal b1-
Man b1-4 Gal b1-4 Glc a1-
Glc b1-6 Man a1-3 Gal a1-
Glc a1-2 Man b1-3 Gal b1-
Glc a1-3 Gal a1-2 Man b1-
Gal a1-2 (Glc b1-6) Man a1-
Gal b1-4 (Man a1-3) Glc a1-
Glc b1-2 (Man a1-3) Gal a1-
Gal a1-6 Glc a1-4 Man b1-
Gal b1-2 Man b1-3 Glc a1-
Man b1-3 Glc a1-2 Gal a1-
Man b1-4 Glc a1-6 Gal b1-
Man b1-6 Gal b1-4 Glc a1-
Glc a1-2 Man a1-4 Gal a1-
Glc a1-3 Man b1-3 Gal b1-
Glc a1-4 Gal a1-2 Man b1-
Gal a1-3 (Glc b1-2) Man a1-
Gal b1-4 (Man a1-6) Glc a1-
Glc b1-2 (Man a1-4) Gal a1-
Gal b1-2 Glc a1-4 Man b1-
Gal b1-3 Man b1-3 Glc a1-
Man b1-4 Glc a1-2 Gal a1-
Man b1-6 Glc a1-6 Gal b1-
Man a1-2 Gal b1-6 Glc a1-
Glc a1-3 Man a1-4 Gal a1-
Glc a1-4 Man b1-3 Gal b1-
Glc a1-6 Gal a1-2 Man b1-
Gal a1-3 (Glc b1-4) Man a1-
Gal b1-6 (Man a1-2) Glc a1-
Glc b1-2 (Man a1-6) Gal a1-
Gal b1-3 Glc a1-4 Man b1-
Gal b1-4 Man b1-3 Glc a1-
Man b1-6 Glc a1-2 Gal a1-
Man a1-2 Glc b1-2 Gal b1-
Man a1-3 Gal b1-6 Glc a1-
Glc a1-4 Man a1-4 Gal a1-
Glc a1-6 Man b1-3 Gal b1-
Glc b1-2 Gal a1-2 Man b1-
Gal a1-3 (Glc b1-6) Man a1-
Gal b1-6 (Man a1-3) Glc a1-
Glc b1-3 (Man a1-2) Gal a1-
Gal b1-4 Glc a1-4 Man b1-
Gal b1-6 Man b1-3 Glc a1-
Man a1-2 Glc a1-3 Gal a1-
Man a1-3 Glc b1-2 Gal b1-
Man a1-4 Gal b1-6 Glc a1-
Glc a1-6 Man a1-4 Gal a1-
Glc b1-2 Man b1-3 Gal b1-
Glc b1-3 Gal a1-2 Man b1-
Gal a1-4 (Glc b1-2) Man a1-
Gal b1-6 (Man a1-4) Glc a1-
Glc b1-3 (Man a1-4) Gal a1-
Gal b1-6 Glc a1-4 Man b1-
Gal a1-2 Man b1-4 Glc a1-
Man a1-3 Glc a1-3 Gal a1-
Man a1-4 Glc b1-2 Gal b1-
Man a1-6 Gal b1-6 Glc a1-
Glc b1-2 Man a1-4 Gal a1-
Glc b1-3 Man b1-3 Gal b1-
Glc b1-4 Gal a1-2 Man b1-
Gal a1-4 (Glc b1-3) Man a1-
Gal a1-2 (Man b1-3) Glc a1-
Glc b1-3 (Man a1-6) Gal a1-
Gal a1-2 Glc a1-6 Man b1-
Gal a1-3 Man b1-4 Glc a1-
Man a1-4 Glc a1-3 Gal a1-
Man a1-6 Glc b1-2 Gal b1-
Man b1-2 Gal b1-6 Glc a1-
Glc b1-3 Man a1-4 Gal a1-
Glc b1-4 Man b1-3 Gal b1-
Glc b1-6 Gal a1-2 Man b1-
Gal a1-4 (Glc b1-6) Man a1-
Gal a1-2 (Man b1-4) Glc a1-
Glc b1-4 (Man a1-2) Gal a1-
Man a1-4 Gal a1-3 Glc b1
Man a1-6 Gal a1-3 Glc b1
Man b1-2 Gal a1-3 Glc b1
Man b1-3 Gal a1-3 Glc b1
Man b1-4 Gal a1-3 Glc b1
Man b1-6 Gal a1-3 Glc b1
Man a1-2 Gal a1-4 Glc b1
Man a1-3 Gal a1-4 Glc b1
Man a1-4 Gal a1-4 Glc b1
Man a1-6 Gal a1-4 Glc b1
Man b1-2 Gal a1-4 Glc b1
Man b1-3 Gal a1-4 Glc b1
Man b1-4 Gal a1-4 Glc b1
Man b1-6 Gal a1-4 Glc b1
Man a1-2 Gal a1-6 Glc b1
Man a1-3 Gal a1-6 Glc b1

31. Major Glycoconjugates of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

32. Major Glycoconjugates of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

33. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

34. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

35. “N-linked” glycan linkage to protein asparagine residues

36. Families of N-linked glycan structures

37. Diversity in complex N-linked glycan structure
Human heart
Human kidney

38. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

39. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

40. “O-linked” glycan linkage to protein serine residues

41. Examples of variations of O-linked glycans

42. Diversity in O-linked glycan structure
Human spleen

43. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

44. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

45. CD59, a complement defense glycoprotein
GPI anchor
plasma membrane

46. GPI (glycosylphosphatidylinositol) anchored protein

47. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

48. Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!

49. Note: No further sugar substitutions needed
O-GlcNAc
Note: No further sugar substitutions needed

50. Three-state model: unmodified / O-GlcNAc / phosphate
Glycoprotein
Unmodified
protein
Phosphoprotein
OPO32-
Phosphatase
OGT
Kinase
O-GlcNAcase
Natasha Zachara

51. Major Glycoconjugates of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

52. Major Glycoconjugates of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

53. Glycolipids are the major glycoconjugates in the brain

54. Examples of brain glycolipids

55. Diversity of brain glycolipids

56. Major Glycoconjugates of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

57. Major Glycoconjugates of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

58. Glycosaminoglycans (GAG’s) and Proteoglycans
long linear glycans made of repeating disaccharides
Hyaluronic acid is the only “stand-alone” GAG, other GAG’s are constituents of …
PROTEOGLYCANS
GAG’s on proteins
Defined by their repeating disaccharide units
GAG’s on proteoglycans are sulfated

59. Essentials of Glycobiology
Hyaluronic acid -- a “simple” space filling molecule
(GlcA β1-3 GlcNAc β1-4)n
Second Edition
Essentials of Glycobiology

60. Heparin / Heparan sulfate
Post-polymerization variations in glycosaminoglycans
Heparin / Heparan sulfate
GlcA β4 GlcNAc α4
IdoA2S α4 GlcNS6S α4

61. Glycosaminoglycan repeating disaccharide units
Second Edition
Essentials of Glycobiology

62. Cartilage proteoglycan

63. Cartilage proteoglycan

64. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions

65. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions

66. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
Biophysical effects
Biochemical effects

67. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
Biophysical effects
Biochemical effects

68. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
General principles
Influenza virus
Inflammation
Biophysical effects
Biochemical effects

69. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
General principles
Influenza virus
Inflammation
Biophysical effects
Biochemical effects

70. Glycans in cell-cell recognition
Carbohydrate recognition domain (CRD)
Carbohydrate binding determinant
Lectin
Glycan
Sharon and Lis (1993) Scientific American

71. Sialoadhesin CRD
May, A.P., et al. (1998) Mol. Cell 1,

72. Principles of glycan-lectin binding:
-hydrogen bonds
-hydrophobic stacking
-multivalent binding
-(salt bridges)
-(calcium coordination)
May, A.P., et al. (1998) Mol. Cell 1,

73. Sialic acids -- Enhanced roles in glycan recognition
glycerol
carboxylate
acetyl
Sialic acid
Comb & Roseman, J Am Chem Soc 80, 497 (1958)

74. Essentials of Glycobiology Second Edition
Diverse sialic acid glycans at the cell surface
sialic acid
Essentials of Glycobiology Second Edition

75. Diverse sialic acid linkages at the cell surface

76. NeuAc-α2,3-Gal-β1,4-GlcNAc

77. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
General principles
Influenza virus
Inflammation
Biophysical effects
Biochemical effects

78. Life cycle of the influenza virus - the role of sialic acids

79. Influenza virus hemagglutinin (side and top views, arrows are sugar binding sites)

80. Sialoglycan structure drives biological recognition: Influenza pandemics are determined by sialic acid linkage specificity
Stevens et al (2006) Nat Rev Microbiol. 4:857-64

81. Influenza virus neuraminidase
(sialidase)

82. Rational design of an anti-influenza drug based on
sialic acid binding to the viral neuraminidase

83. Influenza virus budding in vitro without (top) and with (bottom) neuraminidase inhibitor

84. Prophylactic efficacy of a sialic acid-based influenza neuraminidase inhibitor
Peters, P.H., et al. (2001) J Am Geriatr Soc 49, 1025

85. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
General principles
Influenza virus
Inflammation
Biophysical effects
Biochemical effects

86. Inflammation: How we fight bacterial infection

87. Endothelium
Selectin
Neutrophil
glycoconjugate

88. Sialyl Lewis x (SLex) – NeuAc α2-3 Gal β1-4 (Fuc α1-3) GlcNAc
Leukocyte cell surface glycan ligand required for endothelial recognition
Sialyl Lewis x (SLex) – NeuAc α2-3 Gal β1-4 (Fuc α1-3) GlcNAc
SLex is a minimal binding determinant … a key part of the endogenous selectin ligand

89. Leukocyte adhesion deficiency (LAD) Type II
Marquardt, et al. (1999) J. Pediatr. 134, 681

90. Human blood group antigens and Selectin ligands

91. “Within days of starting fucose therapy, neutrophil levels returned to the normal range .... there were no further infections and antibiotic prophylaxis was discontinued.”
Marquardt, et al. (1999) Blood 94, 3976

92. Pan-selectin ligand GMI-1070
NeuAc-like
Gal-like
Fucose
GlcNAc-like
Tyrosine-sulfate-like
GMI-1070
Completed successful Phase 2 trials for vasoocclusive crisis in sickle cell disease
human Phase 2 results – Telen et al (2015) Blood: March Epub ahead of print human Phase 3 trials –

93. Pan-selectin ligand GMI-1070 blocks leukocyte adhesion-related vaso-occlusive crisis in Sickle Cell Disease (in Phase 2 human trials)
NeuAc-like
Gal-like
GlcNAc-like
Tyrosine-sulfate-like
Fucose
GMI-1070
SLex
mouse model – Chang et al (2010) Blood 116:1779 human Phase 2 trials –

94. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
Biophysical effects
Biochemical effects

95. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
Biophysical effects
Biochemical effects

96. Hyaluronic acid -- a “simple” space filling molecule
(GlcU β4 GlcNAc)n

97. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
Biophysical effects
Biochemical effects

98. Glycobiology Introduction to Glycobiology Glycan structures
Glycan functions
Recognition
Biophysical effects
Biochemical effects

100. Anticoagulant carbohydrate determinant

101. Reference texts Oxford University Press, New York
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Content freely available at:

102. Introduction to Glycobiology
ME:
Course co-directors:
Ronald Schnaar
Natasha Zachara