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Extracellular MacromoleculesExtracellular Macromolecules
Glycosaminoglycans; proteoglycans; Glycosaminoglycans; proteoglycans; glycoproteins; mucinsglycoproteins; mucins
Glycoprotein synthesis; plasma proteinsGlycoprotein synthesis; plasma proteins
Molecular immunology: Molecular immunology: innate immunity; inflammationinnate immunity; inflammation
Molecular immunology: Molecular immunology:
adaptive (acquired) immunityadaptive (acquired) immunity
Fibrous proteins: keratin, collagen and elastinFibrous proteins: keratin, collagen and elastin
Extracellular MacromoleculesExtracellular Macromolecules
1. Glycosaminoglycans Proteoglycans1. Glycosaminoglycans Proteoglycans GlycoproteinsGlycoproteins
MucinsMucins
Extracellular MacromoleculesExtracellular Macromolecules
macromoleculemacromolecule % carb.% carb.
glycosaminoglycansglycosaminoglycans** (GAGs) (GAGs) 100 100
proteoglycansproteoglycans** 90-9590-95
glycoproteinsglycoproteins 2-30 2-30
fibrous proteinsfibrous proteins 1-2 1-2
Examples of functions: Examples of functions:
mechanical supportmechanical support lubricationlubrication
cushioning cushioning adhesivesadhesives
cell spacerscell spacers selective filtersselective filters
* aka* aka mucopolysaccharides, mucoproteins, respectively mucopolysaccharides, mucoproteins, respectively11
Extracellular matrix in tissues Extracellular matrix in tissues ground substance + fibersground substance + fibersmacromolecules between cells macromolecules between cells
ground substance moleculesground substance molecules
GAGs/proteoglycans (mostly carbohydrate)GAGs/proteoglycans (mostly carbohydrate)–fibersfibers
fibrous proteins:fibrous proteins: structural structural
adhesive adhesive
•especially abundantespecially abundantin connective tissuein connective tissue
adhesionmolecules
Adapted from Hypercell
extra-cellularmatrix
basallamina
underlying cells
epithelial cells
2
GAG structureGAG structureexist as: exist as:
independent moleculesindependent moleculese.g.e.g., hyaluronate & heparin, hyaluronate & heparin
parts of larger structuresparts of larger structurese.g.e.g., in proteoglycans, in proteoglycans
heteropolysaccharidesheteropolysaccharides repeating structure: repeating structure:
disaccharide ( disaccharide (ABAB))nn ABABABABABAB…… where where AA is usually 1 uronic acid (hexose with is usually 1 uronic acid (hexose with C6C6 as COO as COO– – )) & & BB is 1 glycosamine (amino sugar) derivative is 1 glycosamine (amino sugar) derivative
unbranchedunbranched glycosidic linkageglycosidic linkage anomeric anomeric CC of 1 unit linked to hydroxyl of adjacent unit of 1 unit linked to hydroxyl of adjacent unit
A sugar
B sugar
33
GAG structure: repeating unitsGAG structure: repeating units GAGGAG A A sugarsugar B sugar B sugarhyaluronatehyaluronate glucuronateglucuronate N-acetyl glucosamineN-acetyl glucosamine
25*
44
GAG structure: repeating unitsGAG structure: repeating units GAGGAG A A sugarsugar B sugar B sugarhyaluronatehyaluronate glucuronateglucuronate N-acetyl glucosamineN-acetyl glucosamine
chondroitin sulfatechondroitin sulfate glucuronate glucuronate N-Ac galactosamine 4-SON-Ac galactosamine 4-SO44
dermatan sulfatedermatan sulfate iduronateiduronate " "
heparan sulfateheparan sulfate glucuronateglucuronate glucosamine N-SOglucosamine N-SO33, 6-SO, 6-SO44
heparin heparin iduronate 2-SOiduronate 2-SO44 " "
keratan sulfatekeratan sulfate galactosegalactose N-Ac glucosamine 6-SON-Ac glucosamine 6-SO44
**opposite configuration in iduronate opposite configuration in iduronate glucuronate/iduronate: glucuronate/iduronate: epimers epimers at at C5C5 glucose/galactose: glucose/galactose: epimersepimers at at C4C4
25*
44
Hyaluronate (aka hyaluronan)mol wt: 106 – 107 (5000 – 50,000 monosaccharide units)very polar: 2 hydroxyls/unit 6 heteroatoms/unit
COO– every other unitbinds cations: Na+, Ca++
213 4 5 6
–
––
(glucuronate–N-acetyl glucosamine)3 (glcUA–glcNAc)3
A B A B A B
Display of HA in motion
55
Hyaluronate: structure & propertiesextended structure (charge repulsion)hydrophilic: binds 10 –100 × wt in H2Oadditional, loosely associated H2O, so that volume occupied ~1000 ×
weight
21 3 4
5 6
–
––
(glcUA–glcNAc)3 glcUAs in space-filling form (CPK)
Display of HA with glcUAs in CPK
66
Hyaluronatesolutions viscous, gel–like, compression-resistant
occurrence: EC matrix,esp. in developing tissue
healing wounds synovial fluid
functions: lubricantshock absorberflexible cementattachment sitepath for cell migration
made by fibroblastsdegraded by hyaluronidase
hyaluronidasebacterial hyaluronidase facilitates spread of infection
Alberts et al. Fig. 19-37
7
HeparinHeparin
mol wt ~ 10mol wt ~ 1044
~ 40 monosaccharide units~ 40 monosaccharide unitsmade & released from mast cells in lungs & livermade & released from mast cells in lungs & liver
88
heparin
cell
HeparinHeparin
mol wt ~ 10mol wt ~ 1044
~ 40 monosaccharide units~ 40 monosaccharide unitsmade & released from mast cells in lungs & livermade & released from mast cells in lungs & liveralso associated with luminal surface of endotheliumalso associated with luminal surface of endotheliumanticoagulantanticoagulant
forms complex with antithrombin IIIforms complex with antithrombin III this complex binds to thrombin, inactivating itthis complex binds to thrombin, inactivating it as a result, clot growth is limitedas a result, clot growth is limited fast-acting, making it therapeutically usefulfast-acting, making it therapeutically useful
88
heparin
cell
Extracellular MacromoleculesExtracellular Macromolecules
macromoleculemacromolecule % carb.% carb.
glycosaminoglycans* (GAGs)glycosaminoglycans* (GAGs) 100 100
proteoglycans*proteoglycans* 90-9590-95
glycoproteinsglycoproteins 2-30 2-30
fibrous proteinsfibrous proteins 1-2 1-2
Examples of functions: Examples of functions:
mechanical supportmechanical support lubricationlubrication
cushioning cushioning adhesivesadhesives
cell spacerscell spacers selective filtersselective filters
* aka* aka mucopolysaccharides, mucoproteins, respectively mucopolysaccharides, mucoproteins, respectively
Proteoglycans (PGs)Proteoglycans (PGs)composed of as many as 200 GAG chains covalently bonded to a core protein via serine side composed of as many as 200 GAG chains covalently bonded to a core protein via serine side
chains chains molecular weight range: 10molecular weight range: 1055 – 10 – 1077 GAG chains:GAG chains: chondroitin sulfate, heparan sulfate, chondroitin sulfate, heparan sulfate, dermatan sulfate, dermatan sulfate,
keratan sulfatekeratan sulfateExamplesExamplesdecorindecorin
many connective tissuesmany connective tissues binds type I collagen, TGF-binds type I collagen, TGF-ββ
perlecanperlecan basal laminaebasal laminae structural & filtering functionstructural & filtering function
aggrecanaggrecansyndecan (syndecan (slide 13slide 13)) GAG chains
coreprotein
99
PG in basal lamina of renal glomerulus
network offibrousproteins &perlecanPG forms filter
adapted from Alberts et al., 3 ed., Fig. 19-56
type IV collagen
laminin
perlecan
entactin
1010
Proteoglycans: aggrecanProteoglycans: aggrecan~100 GAG chains/molecule~100 GAG chains/molecule~100 monosaccharides/GAG chain~100 monosaccharides/GAG chaineach "bristle" = 1 GAG chaineach "bristle" = 1 GAG chaineach GAG chain is either each GAG chain is either chondroitin sulfatechondroitin sulfate
or or keratan sulfatekeratan sulfateGAG chains linked to GAG chains linked to serser side chains of core protein side chains of core protein
corecoreproteinprotein
GAG chainsGAG chains1111
An aggregate of aggrecans & hyaluronanmajor GAG–PG
in cartilagelink proteins bind
noncovalentlywith bound H2O,
disperses shocks,compressive force
~ cell sizeadhesion proteins
link to collagen & cells
degraded by chondroitin sulfatase, etc
ç 1μm è
hyalur-onan
link proteins
keratansulfate chondroitin
sulfate
Alberts et al. Fig. 19-41
core protein
12
Proteoglycans: Proteoglycans: syndecansyndecan
GAG chainsGAG chains
corecoreproteinprotein
cell-surface PGcell-surface PGcore protein domainscore protein domains intracellularintracellular transmembranetransmembrane extracellularextracellular
5 GAGs attached5 GAGs attachedfunctionsfunctions interactionsinteractions
cell-cellcell-cellcell-matrixcell-matrix
growth factor receptorgrowth factor receptorLehninger et al. Fig. 9-22
outside
inside
1313
GAG synthesis & breakdownGAG synthesis & breakdown
synthesissynthesis activated precursors:activated precursors: UDP–monosaccharide UDP–monosaccharide
derivativesderivativese.ge.g., UDP–glucuronate., UDP–glucuronate
residues added one at a time in Golgi complexresidues added one at a time in Golgi complex sulfate moietiessulfate moieties
donor: PAPS (active sulfate)donor: PAPS (active sulfate)degradationdegradation
lysosomeslysosomes specific glycosidases & sulfatasesspecific glycosidases & sulfatases mucopolysaccharidosesmucopolysaccharidoses
genetic disordersgenetic disordersaccumulation of GAG due to absence of a specific glycosidase accumulation of GAG due to absence of a specific glycosidase
or sulfataseor sulfatase
–UDP
1414
adenine––
––
Extracellular MacromoleculesExtracellular Macromolecules
macromoleculemacromolecule % carb.% carb.
glycosaminoglycans (GAGs)glycosaminoglycans (GAGs) 100 100
proteoglycansproteoglycans 90-9590-95
glycoproteins*glycoproteins* 2-30 2-30
fibrous proteinsfibrous proteins 1-2 1-2
* polypeptide with 1 or more oligosaccharide side chains* polypeptide with 1 or more oligosaccharide side chains
1515
Glycoproteins: functions of glyco moietiesGlycoproteins: functions of glyco moietiesincrease protein’s solubility & hydrophilicity (increase protein’s solubility & hydrophilicity (sl 19sl 19))stabilize protein againststabilize protein against
denaturationdenaturation proteolysisproteolysis
markersmarkers direct protein's destinationdirect protein's destination
organelleorganelleplasma membraneplasma membraneexport (secretion)export (secretion)
indicate protein's lifetime (indicate protein's lifetime (sl 21sl 21)) part of the protein's receptor recognition site (part of the protein's receptor recognition site (sl 23sl 23))
signal factors such as hormones, cytokinessignal factors such as hormones, cytokinescell-cell adhesion proteinscell-cell adhesion proteins
Glycosylation:Glycosylation: one kind of one kind of post-translational modificationpost-translational modificationothers:others: phosphorylation phosphorylation
carboxylation carboxylation
1616
Glycoprotein structureGlycoprotein structurepolypeptide with 1 or more oligosaccharide side chainspolypeptide with 1 or more oligosaccharide side chainsoligosaccharide linked to polypeptide in two ways:oligosaccharide linked to polypeptide in two ways:typetype linked to side chain of linked to side chain of organelle where sugarsorganelle where sugars
are added to proteinare added to protein
O-linkedO-linked serine (serine (serser), threonine (), threonine (thrthr), ), Golgi complex lumenGolgi complex lumen (O-glycoside)(O-glycoside) hydroxylysine (in collagen)hydroxylysine (in collagen)
N-linkedN-linked asparagine (asparagine (asnasn)) rough ER lumenrough ER lumen (N-glycoside)(N-glycoside)
1717
Glyco moiety structureoligosaccharide chain extends away from protein surfaceoligosaccharide chain extends away from protein surfaceunits mostly hexoses in pyranose (6-atom ring) formunits mostly hexoses in pyranose (6-atom ring) formbranchedbranchedglycosidic links varied: glycosidic links varied:
αα or or ββ1,2; 1,3; 1,41,2; 1,3; 1,4
terminal terminal sugarsugaroften often sialatesialate
asn
2
7
2
7
Stryer 4ed., p. 4631818
asn
Mucins: salivary glycoproteinsMucins: salivary glycoproteins
mol wt ~ 10mol wt ~ 1066
~800 short ~800 short (disaccharide) (disaccharide) side chainsside chains
terminal sugar is terminal sugar is sialatesialate anionic sugaranionic sugar at end of glyco chains at end of glyco chains
of many glycoproteinsof many glycoproteinsvery hydrophilic, very hydrophilic,
extended structureextended structure
galNAcsialate
–
2
~ ~
1919
Mucins: modification & aggregationMucins: modification & aggregation
sialidase sialidase (neuraminidase)(neuraminidase) catalyzes hydrolysiscatalyzes hydrolysis
of sialates from mucins of sialates from mucins secreted by oral bacteriasecreted by oral bacteria
products:products: less hydrophilic, less hydrophilic,
less Hless H22O-soluble, O-soluble, more folded, more folded, more aggregatedmore aggregated
part of the enamel pellicle part of the enamel pellicle & dental plaque matrix& dental plaque matrix
sialidasex H2O galNAc
sialate
~ ~
~~
x
~~
2020
Role of glyco moiety in controlling protein lifetimeRole of glyco moiety in controlling protein lifetime
many blood proteins have glyco chains with terminal many blood proteins have glyco chains with terminal sialatesialateendothelial surface endothelial surface sialidasessialidases slowly remove sialates from these circulating proteins slowly remove sialates from these circulating proteinsrate of sialate removal depends on protein's structurerate of sialate removal depends on protein's structure
now-exposed now-exposed gal–glcNAcgal–glcNAc…… residues bind to residues bind to asialoglycoprotein receptorasialoglycoprotein receptor on liver cell on liver cell surfacesurface
protein is then endocytosed & broken downprotein is then endocytosed & broken down
sialoglycoprotein: sia–gal–glcNAc–[core sugars]–protein
asialoglycoprotein: gal–glcNAc–[core sugars]–protein
2121
Blood group typesBlood group types
A: have A: have –– enzyme to add enzyme to add galNAcgalNAc to core sugars to core sugars–– antibody to type B antigen antibody to type B antigen
B: have B: have –– enzyme to add enzyme to add galgal to core sugars to core sugars–– antibody to type A antigen antibody to type A antigen
O: have O: have –– neither enzyme neither enzymeAB: have AB: have –– both enzymes (either both enzymes (either galNAcgalNAc or or galgal
added to core sugars)added to core sugars)
Type O cell surface: gal–glcNAc–gal–glc–protein†
Type A cell surface: galNAc–gal–glcNAc–gal–glc–protein†
Type B cell surface: gal–gal–glcNAc–gal–glc–protein†
* 6-deoxygalactose†or lipid
both antibodies
neither antibody
|fucose
|fucose
|fucose*
2222
core sugarscore sugars
Glyco moiety-binding proteins: lectinsGlyco moiety-binding proteins: lectinscontain sites that bind specific glyco structurescontain sites that bind specific glyco structurese.g., e.g., asialoglycoprotein receptorasialoglycoprotein receptor described on described on sl 21sl 21important in intercell adhesion (i.e., lectins are CAMs: important in intercell adhesion (i.e., lectins are CAMs: cell cell
adhesion moleculesadhesion molecules))selectins:selectins:
plasmaplasmamembranemembranelectins thatlectins thatmediatemediatecell-cellcell-cellrecognitionrecognition& adhesion& adhesion
Lehninger et al. Fig. 7-372323