Download presentation
Presentation is loading. Please wait.
1
Introduction to glycobiology
Course Objectives Learn the basic language of glycobiology Monosaccharides (vertebrate) Glycans (oligosaccharide & glycoconjugate basic concepts) Learn basic glycan functions Glycans in glycoprotein activity, folding, trafficking O-GlcNAc-mediated regulation Glycolipids in membrane recognition & regulation Proteoglycans in the extracellular matrix Glycan binding proteins Learn the basic concepts of glycan biosynthesis Learn the basic tools of glycomics (analysis)
2
Introduction to glycobiology
Date Day Lecture Faculty ONLINE Saccharides & The Glyco World Dr. R. Schnaar Mar 19 Wed Glycoproteins I Dr. N. Zachara Mar 21 Fri Glycoproteins II Mar 25 Tue Glycolipids and GPI anchors Mar 27 Thu Carbohydrate Engineering Dr. K. Yarema Mar 31 Mon O-GlcNAc Dr. G. Hart Apr 2 Glycomics (Analytical Glycobiology) Apr 4 Glycans and Disease Apr 8 Hyaluronan and Proteoglycans Apr 10 Protein-Glycan Recognition (Lectins) Apr 14 Glycan Binding Protein Functions
3
Introduction to glycobiology
Course co-directors: Ronald Schnaar Natasha Zachara Lecturers: Gerald Hart Kevin Yarema
4
Introduction to glycobiology
Oxford University Press, New York Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Content freely available at:
5
What are: Glycobiology? Glycosciences? Glycomics?
6
Glycobiology The study of the biological functions, structures, recognition and biosynthesis of glycans (sugar chains, saccharides) in the context of the biological scaffolds to which they are attached (e.g. glycolipids & glycoproteins).
7
Glycosciences Glycobiology & Glycochemistry, including…
-Chemical synthesis -Chemoenzymatic synthesis -Metabolic engineering -Glycomimetics -Glycan conformation and structure -Material Sciences -Plant and bacterial polysaccharides
8
Glycomics - Analytical glycosciences -Glycan bioinformatics
“Glycome” – The total complement of glycans in a cell or organism
9
NOT Carbohydrates as food
Glycobiology is … NOT Carbohydrates as food Carbohydrates in food are important sources of energy. Starch found in plant-derived food such as pasta .... Berg, Tymoczko & Stryer (2006) Biochemistry, Sixth Edition, W.H. Freeman, New York
10
Glycobiology is NOT “a” post-translational modification
Glycobiology is NOT “a” post-translational modification... it is thousands Combinatorial linkage position, orientation and branching provide the potential for millions* of different glycan structures – functional diversity Glycans may be larger and are more diverse than their (protein) carriers A glycan’s function can supersede that of its (protein) carrier 1-2% of the human genome is devoted to glycosylation Modified from faculty/hougland.html Cummings RD (2009) Molecular BioSystems 5, 1087 *Glycoproteins and glycolipids may contain ~3000 glycan determinants with an additional ~4000 theoretical pentasaccharide sequences in glycosaminoglycans
11
Glycobiology is … NOT a “decoration”
N-linked carbohydrate can form both an immunologically silent face—with carbohydrate masquerading as "self"—and also can protect neighboring epitopes through an "evolving glycan shield" structuralbiologylaboratory/Pages/kwong.aspx “Structure” of HIV gp120 Zolla-Pazner (2004) Nature Reviews Immunology 4, 199
12
CD59, a complement defense glycoprotein
Proteins typically fold inward, whereas glycans spread out in space 3 kDa N-linked glycan 20 kDa protein 1 kDa O-linked glycan “naked” CD59, Huang et al (2007) Acta Crystallographica 63, 714 1.5 kDa GPI anchor CD59, a complement defense glycoprotein Rudd et al (1997) J Biol Chem 272, 7229
13
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
14
Unique to yeast – NO! 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
15
The Eukaryotic Cell Surface
? Tiny sugar “decorations”.. added as an afterthought Lodish, et al. (1995) Molecular Cell Biology 3rd ed.
16
The Cell Surface -- The Real Picture
extracellular lipid bilayer fibroblast intracellular Martinez-Palomo, A., et al. Cancer Res. 29, , 1969 The “glycocalyx” surrounding a fibroblast. Cell surface carbohydrates are stained black.
17
Tropical Forest Canopy
“Evolution has failed to generate a living cell devoid of surface glycosylation” - A. Varki Cohen & Varki (2010) OMICS 4:455
18
Real image textbook image
The Cell Surface Real image textbook image lipid bilayer fibroblast intracellular Martinez-Palomo, A., et al. Cancer Res. 29, , 1969 Among their diverse functions, glycans … encode intermolecular and cell-cell recognition regulate the activities of proteins (enzymes, ion channels, receptors) Define biophysical space (large, hydrated)
19
Glycobiology = Language (semiotics)
Cohen & Varki (2010) OMICS 4:455 Rosetta Stone, British Museum
20
Saccharides Carbohydrates: The language of glycobiology
The glycosidic bond and glycan nomenclature Major vertebrate glycan classes
21
Saccharides Carbohydrates: The language of glycobiology
The glycosidic bond and glycan nomenclature Major vertebrate glycan classes
22
(CH2O)n = “carbo” “hydrate”
Carbohydrates – the building blocks of Glycobiology (CH2O)n = “carbo” “hydrate” Generalized Ketose Generalized Aldose
23
glucose fructose
24
Monosaccharide structural identity is all about stereochemistry
Generalized Aldose Monosaccharide structural identity is all about stereochemistry Enantiomers: mirror images of each other that are not superimposable. Diastereomers: stereoisomers that are not enantiomers.
25
Fischer projection D-glyceraldehyde
26
Enantiomers * * D-glyceraldehyde L-glyceraldehyde *
Highest numbered asymmetric carbon = reference carbon
27
Enantiomers * * D-glyceraldehyde L-glyceraldehyde *
Highest numbered asymmetric carbon = reference carbon
28
Diasteriomers * * * Highest numbered asymmetric carbon = reference carbon
29
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
30
Essentials of Glycobiology
* * epimers: differ in only stereogenic center Essentials of Glycobiology Second Edition
31
Monosaccharides equilibrate between open chain and ring forms
Monosaccharides equilibrate between open chain and ring forms. Two ring forms (anomeric configurations) are possible, α and β α-D-glucose β-D-glucose D-glucose Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition
32
Determining anomeric configuration
[PDB] Lee & Lee (1999) J Chinese Chem Soc 46:
33
Monosaccharide conformation – Chair configurations
Taylor & Drickamer (2006) Introduction to Glycobiology, 2nd edition
34
Nearly all vertebrate glycans are built from only 9 sugars:
3 hexoses* Glucose Mannose Galactose 2 N-acetylhexosamines N-acetylglucosamine N-acetylgalactosamine xylose glucuronic acid sialic acid L-fucose *all D configuration except fucose
35
Seven eukaryotic sugars and their relationship to glucose
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition
36
Two important terminal eukaryotic sugars
Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition
37
Glycan properties for molecular recognition and binding energy
Glucose Galactose Mannose Sialic Acid
38
Saccharides Carbohydrates: The language of glycobiology
The glycosidic bond and glycan nomenclature Major vertebrate glycan classes
39
Saccharides Carbohydrates: The language of glycobiology
The glycosidic bond and glycan nomenclature Major vertebrate glycan classes
40
Glycosidic bonds link sugars together
(lactose) CCRC When a glycosidic bond is formed the anomeric configuration is “locked” Taylor & Drickamer (2011) Introduction to Glycobiology, 3rd edition
41
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
42
Nomenclature: Branches are placed in parentheses
NeuAc α2-3 Gal β1-4 (Fuc α1-3) GlcNAc
43
Varki et al. (2009) Proteomics. 9:5398
44
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
45
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 …
46
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 Cummings RD (2009) Molecular BioSystems 5, 1087 *Glycoproteins and glycolipids may contain ~3000 glycan determinants with an additional ~4000 theoretical pentasaccharide sequences in glycosaminoglycans
47
Saccharides Carbohydrates: The language of glycobiology
The glycosidic bond and glycan nomenclature Major vertebrate glycan classes
48
Saccharides Carbohydrates: The language of glycobiology
The glycosidic bond and glycan nomenclature Major vertebrate glycan classes
49
Major Glycans of Eukaryotes
Glycoproteins Glycolipids Proteoglycans
50
Major Glycans of Eukaryotes
Glycoproteins Glycolipids Proteoglycans
51
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
52
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
53
“N-linked” glycan linkage to protein asparagine residues
54
Families of N-linked glycan structures
55
Terminal Saccharide Diversity --- A key to recognition
Examples of structures found as termini on the branches of complex N-linked glycoprotein glycans…. NeuAc 2-6 Gal 1-4 GlcNAc--- NeuAc 2-3 Gal 1-4 GlcNAc--- Gal 1-3 Gal 1-4 GlcNAc--- 4-SO3-GalNAc 1-4 GlcNAc--- (-Gal 1-4 GlcNAc 1-3-)n = polylactosamine repeats (n may be >6) (-NeuAc 2-8-) n = polysialic acid (n may be >50) Fuc residues (e.g. Fuc 1-2 Gal; Fuc 1-3 GlcNAc; Fuc 1-4 GlcNAc
56
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
57
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
58
“O-linked” glycan linkage to protein serine residues
59
Examples of variations of O-linked glycans
60
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
61
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
62
GPI (glycosylphosphatidylinositol) anchored protein
63
CD59, a complement defense glycoprotein
3 kDa N-linked glycan 20 kDa protein 1 kDa O-linked glycan 1.5 kDa GPI anchor plasma membrane
64
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
65
Glycoproteins N-Linked O-Linked GPI Anchor O-GlcNAc
*A non-inclusive list!
66
O-GlcNAc Note: No further sugar substitutions have been confirmed
Highly dynamic post-translational modification!
67
Major Glycans of Eukaryotes
Glycoproteins Glycolipids Proteoglycans
68
Major Glycans of Eukaryotes
Glycoproteins Glycolipids Proteoglycans
69
Galactosylceramide (GalCer)
A Glycosphingolipid Galactosylceramide (GalCer)
70
A Ganglioside “GD1a”
71
Major Glycans of Eukaryotes
Glycoproteins Glycolipids Proteoglycans
72
Major Glycans of Eukaryotes
Glycoproteins Glycolipids Proteoglycans
73
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
74
Hyaluronic acid -- a “simple” space filling molecule
(GlcU β4 GlcNAc)n
75
Post-polymerization variations in glycosaminoglycans
76
Cartilage proteoglycan
77
Cartilage proteoglycan
78
Glycan Biosynthesis Glycosyltransferase reaction (UDP-Gal:glucose β4 galactosyltransferase) UDP-Glc UDP-Gal GDP-Man UDP-GlcNAc UDP-GalNAc UDP-GlcA UDP-Xyl GDP-Fuc CMP-NeuAc Activated Sugars:
79
Glycoprotein biosynthesis
N-Linked – en-bloc preconstructed core, trimming, terminal elaboration O-Linked – stepwise sugar by sugar addition O-GlcNAc - dynamic transferase/glycosidase GPI Anchor – en-bloc preconstructed core, elaboration Glycolipid biosynthesis Stepwise sugar-by-sugar addition Proteoglycan biosynthesis Stepwise sugar-by-sugar addition (core and repeating disaccharide) Post-polymerization modifications
Similar presentations
© 2025 SlidePlayer.com. Inc.
All rights reserved.