1. Glycobiology resources
“Concise, clear introduction”
“Thorough, detailed, comprehensive”
Oxford University Press, New York
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. FREE –
& coming soon:
Schnaar RL “Glycobiology Simplified” J Leuk Biol 99:825

2. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

3. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

4. 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

5. A MAJOR cell component invisible to standard phase microscopy
Light (phase) 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 (phase) micrograph of Cryptococcus neoformans
Steenbergen et al. (2003) Microbes and Infection 5:667
Scanning electron micrograph of C. neoformans yeast cells.
Van Duin et al. (2004) Antimicrobial Agents and Chemotherapy 48:2014

6. Evolution has failed to generate a living cell devoid of surface glycosylation
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

7. Tropical Forest Canopy
Cohen & Varki (2010) OMICS 4:455
Ajit Varki, “Perspectives on the Future of Glycobiology,” Athens, GA, 14-Mar-2011

8. The Cell Surface Textbook image Real image fibroblast
Martinez-Palomo, A., et al. Cancer Res. 29, , 1969

9. Glycobiology = Language (semiotics)
Cohen & Varki (2010) OMICS 4:455
Rosetta Stone, British Museum

10. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

11. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

12. (CH2O)n = “carbo” “hydrate”
Carbohydrates – the building blocks of Glycobiology
(CH2O)n = “carbo” “hydrate”
Generalized Ketose
Generalized Aldose

13. D-glucose
D-fructose

14. mirror images of each other that are not superimposable.
Sugar diversity – structurally and functionally – is all about STEREOCHEMISTRY
Enantiomers:
mirror images of each other that are not superimposable.
In sugar nomenclature, enantiomers get a letter “D” or “L”
Diastereomers:
stereoisomers that are not enantiomers.
In sugar nomenclature, diastereomers get different names

15. Enantiomers Mirror images of each other that are not superimposable.
In sugar nomenclature, enantiomers get a letter “D” or “L” * *
D-glyceraldehyde
L-glyceraldehyde *
Highest numbered asymmetric carbon = reference carbon

16. Fischer projection
D-glyceraldehyde

17. Diasteriomers Stereoisomers that are not enantiomers.
In sugar nomenclature, diastereomers get different names * * *
Highest numbered asymmetric carbon = reference carbon

18. diasteriomers & enantiomers
Enantiomers & Diasteriomers * *
carbons
asymmetric carbons
diaster- iomers
diasteriomers & enantiomers 3 1 2 4 5 8 6 16 * * *
Highest numbered asymmetric carbon = reference carbon

19. Essentials of Glycobiology*
* epimers: differ in one stereogenic center
Essentials of Glycobiology
Second Edition

20. Monosaccharides are predominantly in RING FORM at equilibrium.
Free sugars equilibrate between open chain and ring forms.
Two ring forms (anomeric configurations) are possible, α and β
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition

21. Determining anomeric configuration
Step 1: D or L?
Highest numbered asymeric carbon
Lee & Lee (1999) J Chinese Chem Soc 46:

22. Monosaccharide conformation – Chair configurations
Taylor & Drickamer (2006) Introduction to Glycobiology, 2nd edition

23. Just nine monosaccharides are responsible for building nearly all mammalian glycans.
glucose, galactose, mannose
N-acetylglucosamine, N-acetylgalactosamine
xylose
glucuronic acid
fucose
sialic acid
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.

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

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

26. Glycan properties for molecular recognition and binding energy
Glucose
Galactose
Mannose
Sialic Acid

27. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

28. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

29. Glycosidic bonds link sugars together
(lactose)
When a glycosidic bond is formed the anomeric configuration is “locked”
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition

30. 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
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition

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

32. Varki et al. (2009) Proteomics. 9:5398

33. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

34. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

35. Sugars, polysaccharides, oligosaccharides and glycans
Sugars: often from plants or breakdown products of plant polysaccharides.
Examples: glucose, sucrose (glucose α1-2 fructose), maltose (Glc α1-4 Glc), lactose (Gal β1-4 Glc).
Polysaccharides: long polymers (linear or branched) of a single monosaccharide
Examples: Cellulose, amylose (starch), glycogen, chitin
Oligosaccharides: groups of monosaccharides (typically 3 or more) in linear or branched glycosidic linkage
Seldom found as free structures – components of glycans
Glycans: Oligosaccharides on proteins or lipids
Examples: Glycoproteins, glycolipids

36. Examples of polysaccharides: Cellulose
Most abundant organic compound on Earth. ~90% of cotton fiber, 50% of wood, 33% of all plant matter.
(β1-4 Glc β1-4 Glc)n
Intra- and inter-strand hydrogen bonding provides strength
Material sciences

37. Examples of polysaccharides: Amylose
Linear (amylose) and branched (amylopectin) forms are the carbohydrates of starch. This major food source is enzymatically broken down to glucose during digestion…
(α1-4 Glc α1-4 Glc)n
…and during ethanol production
α1-6 Glc branches

38. Examples of polysaccharides: Glycogen
Tissue storage form of glucose (primarily in the liver & muscle). “Animal Starch” – same general structure as plant starch, but more branched & compact. Broken down to glucose at need to quickly increase blood glucose levels.
(α1-4 Glc α1-4 Glc)n
α1-6 Glc branches

39. Examples of polysaccharides: Chitin
Among the most abundant of organic molecules on Earth, it is the main component of the cell walls of fungi, the exoskeletons of arthropods such as insects and crustaceans (e.g., crabs, lobsters and shrimps).
(β1-4 GlcNAc β1-4 GlcNAc)n

40. oligosaccharides as molecular language
Cohen & Varki (2010) OMICS 4:455
Rosetta Stone, British Museum

41. Oligosaccharides: 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

42. 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 …

43. 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

44. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

45. saccharides, glycosides & glycans
The Glyco World
saccharides, glycosides & glycans
Glycans: the "missing mass" of the biological universe
Carbohydrates: The language of glycoscience
The glycosidic bond and glycan nomenclature
Major carbohydrate classes: sugars, polysaccharides, oligosaccharides and glycans
Major glycans of eukaryotes

46. Major Glycans of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

47. Major Glycans of Eukaryotes
Glycoproteins
Glycolipids
Proteoglycans

48. Glycans are large hydrated surface components of glycoproteins
Burton DR et al (2012) Cell Host & Microbe 4:396
HIV gp120

49. HIV gp120
Doores K J et al. (2010) PNAS 107:17107

50. 3 kDa N-linked glycans 1 kDa N-linked glycan 18 kDa protein
Proteins fold onto themselves… glycans spread out in space
Thy-1 cell-cell interaction protein
1.5 kDa GPI anchor
3 kDa N-linked glycans
18 kDa protein
membrane
Second Edition
Essentials of Glycobiology
1 kDa N-linked glycan

51. Glycoproteins Roles of glycoprotein glycosylation
Modes of glycoprotein glycosylation
N-linked glycans
O-linked glycans
O-GalNAc – “mucin” type glycosylation
O-Man, O-Fuc, O-Glc, O-Xyl – signaling molecules
O-GlcNAc – intracellular protein regulation
Glycan Biosynthesis
General Principles
N-linked glycoprotein biosynthesis

52. Glycoproteins Roles of glycoprotein glycosylation
Modes of glycoprotein glycosylation
N-linked glycans
O-linked glycans
O-GalNAc – “mucin” type glycosylation
O-Man, O-Fuc, O-Glc, O-Xyl – signaling molecules
O-GlcNAc – intracellular protein regulation
Glycan Biosynthesis
General Principles
N-linked glycoprotein biosynthesis

53. Roles of protein glycosylation
Protein conformation
Protein folding*
Protein trafficking*
Cell surface glycoprotein lattice*
Regulation of signaling pathways*
Cell-cell interactions*
* via glycan recognition

54. Protein Conformation:
Closely spaced short O-linked glycans on mucins

55. Protein Folding:
N-linked protein glycosylation is essential for chaperone-assisted folding of glycoproteins in the endoplasmic reticulum
Asn
Helenius, A. and Aebi, M. (2001) Science 291,

56. Chemical Biology: Glycans
Protein Conformation:
10/17/2013
Conserved glycosylation of IgG is required for interaction with immune effectors (complement, Fc-receptors)

57. Protein Trafficking:
Unique glycosylation of lysosomal enzymes “labels” them for trafficking to the lysosome via the Man-P receptor(s)
Asn

58. Cell surface glycoprotein lattice:
Sugar binding proteins (galectins) bind to and stabilize cell surface glycoproteins
Altered glycoprotein glycans result in internalization of key glycoproteins
Pancreatic β cells stained for Glut-2 (green), insulin (red), and DNA (blue). Ohtsubo et al (2005) Cell 123:1307
Wild type
Glycosyaltion mutant
Lau et al (2007) Cell 129:123

59. Regulation of signaling pathways:
Notch signaling during development is driven by glycosylation
Stanley & Okajima (2010) Curr Topics Devel Biol 92:131
Stanley (2007) Curr Opin Struct Biol 17:530

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

61. Roles of protein glycosylation
Protein conformation
Protein folding*
Protein trafficking*
Cell surface glycoprotein lattice*
Regulation of signaling pathways*
Cell-cell interactions*
* via glycan recognition