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Bacterial PolysaccharidesBacterial PolysaccharidesAnimal PathogensAnimal Pathogens•• Salmonella Salmonella enteriditisenteriditis••E. coliE. coli••NeisseriaNeisseria meningitidismeningitidis••HaemophilusHaemophilus influenzaeinfluenzae••Proteus mirabilisProteus mirabilis••Proteus Proteus vulgarisvulgaris••MycobacteriaMycobacteria(many others)(many others)
Soil BacteriaSoil Bacteria•• RhizobiaceaeRhizobiaceae••Pseudomonas Pseudomonas solanacearumsolanacearum••ErwiniaErwinia amylovoraamylovora(many others)(many others)
PolysaccharidesPolysaccharides••ExtracellularExtracellular polysaccharides (polysaccharides (EPSsEPSs))
••Excreted Excreted heteropolysaccharidesheteropolysaccharides••Most often acidicMost often acidic••Frequently form solutions of high viscosityFrequently form solutions of high viscosity••GramGram--positive and Grampositive and Gram--negative productsnegative products
••Capsular Polysaccharides (Capsular Polysaccharides (CPSsCPSs))••CellCell--associated associated heteropolysaccharidesheteropolysaccharides••Most often acidic, but some are neutralMost often acidic, but some are neutral••Solutions can be highly viscousSolutions can be highly viscous••GramGram--positive and Grampositive and Gram--negative productsnegative products
••LipopolysaccharidesLipopolysaccharides ((LPSsLPSs))••Component of the outer membraneComponent of the outer membrane••AmphiphilicAmphiphilic••GramGram--negative bacterial componentnegative bacterial component
••TeichoicTeichoic acidacid••GramGram--PositivePositive
••Mycobacterium cell wall componentsMycobacterium cell wall components••MycolylMycolyl arabinogalactanarabinogalactan••LipooligosaccharidesLipooligosaccharides ((LOSsLOSs))••GlycopeptidolipidsGlycopeptidolipids ((GPLsGPLs))••PhenolicPhenolic glycolipidsglycolipids ((PGLsPGLs))
Polysaccharide FunctionsPolysaccharide Functions
••Prevention of Prevention of desicationdesication••SurvivialSurvivial
••AdherenceAdherence••Colonization of oral surfacesColonization of oral surfaces••Colonization of cathetersColonization of catheters••BacteriaBacteria--plant interactionsplant interactions
••Resistance to nonResistance to non--specific host immunityspecific host immunity••ComplementComplement--mediated mediated phagocytosisphagocytosis••ComplementComplement--mediated killingmediated killing
••Resistance to specific host immunityResistance to specific host immunity••Poor antibody responsePoor antibody response
••CellCell--cell recognition cell recognition signallingsignalling••BacteriaBacteria--plant interactionsplant interactions
GramGram--Negative Bacterial Cell WallNegative Bacterial Cell Wall
CPS
EPS
E. coliE. coli Capsular PolysaccharideCapsular Polysaccharide
E. coli K30 wt
E. coli K30CPS minus
E. coli K30CPS/LPS minus
From"Current Topics in Microbiology and Immunology", 1990, Ed. by K. Jann and B. Jann
Examples of Capsular Polysaccharides fromExamples of Capsular Polysaccharides fromEnteric BacteriaEnteric Bacteria
44--ββ--DD--GlcAGlcA--1 41 4--αα--DD--GlcNAcGlcNAc--1 61 6--αα--DD--GlcNAcGlcNAc--11 nnLL--SerineSerine--6 6 (amide)(amide)
E. coliE. coli K40 CPSK40 CPS
22--αα--DD--ManMan--1 31 3--ββ--DD--GalGal--11 nnββ--DD--GlcAGlcA--1 31 3--αα--DD--GlcGlc--33
E. coliE. coli K30 CPSK30 CPS
Grou
p I
Grou
p I
Grou
p II
Grou
p II 88--αα--Neu5AcNeu5Ac--2 92 9--αα--Neu5AcNeu5Ac--22 nn
E. coliE. coli K92 CPSK92 CPS
33--ββ--DD--RibRib--1 71 7--ββ--KdoKdo--22 nnE. coliE. coli K23 CPSK23 CPS
(Whitfield and Valvano. 1993. Biosynthesis and expression(Whitfield and Valvano. 1993. Biosynthesis and expressionof cellof cell--surface polysaccharides in gramsurface polysaccharides in gram--negative bacteria. negative bacteria. Adv. Microbial. Adv. Microbial. PhysiolPhysiol..35:13535:135--246).246).
LipopolysaccharidesLipopolysaccharides: Structure, Synthesis, : Structure, Synthesis, FunctionFunction
READ: Raetz, and Whitfield. 2002. READ: Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Schematic Diagram of a Schematic Diagram of a LipopolysaccharideLipopolysaccharide
Lipid ACore
Lipid ACoreO-Chain
Lipid ACoreO-Chain
LPS II
LPS I
LPS III, IV, V
Lipid A BiosynthesisLipid A Biosynthesis
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Lipid A and the Innate Immune ResponseLipid A and the Innate Immune Response
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Unusual Lipid A StructuresUnusual Lipid A Structures
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
RhizobialRhizobial and and E. coliE. coli Lipid ALipid A
Rhizobiaceae
R. leguminosarumR. etli
O
OHHO
HO CO2 HCore
OHO
O
O
NHO
O O
O NHCO2 H
OHHO
CH2 O
OHO
HOOO
O
O
HO O
Rhizobium Lipid AE. coli Lipid A
(U.R. Bhat, L.S. Forsberg, & R.W. Carlson. 1994.J. Biol. Chem. 269:14402-14410; N. L. Que, S. Lin, R.J.Cotter & C.R. Raetz.2000. J. Biol. Chem. 275:28006-28016; N.L. Que, A.A. Ribeiro & C.R. Raetz2000. J. Biol. Chem. 275:28017-28027)
OO
O
NHO
O OHO
ONH
O
O
P
P
O
O
HO
OH
OH
OH
OHO
O
HO
O
O
O O
HO
((BrucellaBrucella, , BartonellaBartonella))
Biosynthesis of Unusual Lipid A StructuresBiosynthesis of Unusual Lipid A Structures
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
O
NHO
-O3POO
O
HO
O
NHO
HOOPO3
-
O
HO CH2OH
CO2H
HO
O
O
HO
HO CH2OH
CO2H
HO
O
O
HOO
HO
O
HO
UDP-GlcNAc1
1. 3-O-acylation (lpxA).2. De-N-acetylation (lpxC).3. N-acylation (lpxD).4. Disaccharide synthase (lpxB).5. 4’-phosphorylation (kinase).6. Kdo transferase (kdtA).
1. Price, Kelly, Raetz, and Carlson. 1994. J. Bacteriol.176:4646-4655.
2. Price, Jeyaretnam, Carlson, Kadrmas, and Raetz. 1995.PNAS 92:7352-7356.
3. Brozek, Kadrmas, and Raetz. 1996. JBC 271:32112-32118.
1-phosphatase34’-phosphatase2
27O
HC
28:0
Acp
-XL
4
4. Brozek, Carlson, and Raetz. 1996. JBC 271:32126-32136.
Man
LpcC5
5. Kadrmas, Allaway, Studholme, Sullivan, Ronson, Poole, and Raetz. 1998. JBC 273:26432-26440.
GalLpcA5
KdoLpcB5
GalA ?
GalA ?
GalA ?
GalA?
Glc
Nox
idat
ion7
(?)
acyl
ase6
6. Basu, White, Que, and Raetz. 1999. JBC 274:11150-11158.
7. N. L. S. Que, S. H. Lin, R. J. Cotter, and C. R. H. Raetz. 2000.JBC 275 (36):28006-28016.
Biosynthesis ofRhizobial LPS
Transport of Lipid ATransport of Lipid A
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Transport of Lipid ATransport of Lipid A
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Lipid A Core StructuresLipid A Core Structures
Schematic Diagram of a Schematic Diagram of a LipopolysaccharideLipopolysaccharide
Lipid ACore
Lipid ACoreO-Chain
Lipid ACoreO-Chain
LPS II
LPS I
LPS III, IV, V
Lipid A Core StructuresLipid A Core Structures
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Core BiosynthesisCore Biosynthesis
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Polymerization and Polymerization and LigationLigation of Oof O--Chain PolysaccharideChain Polysaccharide
n
0123456
Pare
nt
rfc
mut
ant
rfaL
mut
ant
Num
ber o
f rep
eat u
nits
(Figure taken from Whitfield. 1995. Biosynthesis of (Figure taken from Whitfield. 1995. Biosynthesis of lipopolysaccharidelipopolysaccharide O antigens.O antigens.Trends in MicrobiologyTrends in Microbiology 3:1783:178--185)185)
LipopolysaccharidesLipopolysaccharides: Structure, Synthesis, : Structure, Synthesis, FunctionFunction
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Rhizobium leguminosarum/etli LPS Core
O-Chain
O
O
CO 2H
O
CH 2O HO H
Lipid A
CO 2H
H O CH 2O H
OO
CO 2H
O
O
O
O
CO 2H
O
O
CO 2H
OH O
CH 2O HHO
O CO 2H
O
O
O
O H
H O
O H
O H
O H
O H
O H
O H
O H
H O
H O
H O
H O
O H
Kdo
Gal
Man
KdoGalA
GalA
GalA
Kdo
(U.R. Bhat, S.K. Bhagyalakshmi, & R.W. Carlson. 1991. Carbohydr. Res. 220:219-227,L.S. Forsgberg, & R.W. Carlson. 1998. J. Biol. Chem. 273:2747-2757)
LipopolysaccharidesLipopolysaccharides: Structure, Synthesis, : Structure, Synthesis, FunctionFunction
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
LipopolysaccharidesLipopolysaccharides: Structure, Synthesis, : Structure, Synthesis, FunctionFunction
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
LipopolysaccharidesLipopolysaccharides: Structure, Synthesis, : Structure, Synthesis, FunctionFunction
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.
Regulation of Polysaccharide Chain LengthRegulation of Polysaccharide Chain Length
rfb
rol
rfb
rol
rfb
rol
+
+
+
1 kb1 kb3 kb
orf1 orf2 orf3 wzm wzt
FAJ1
201
FAJ1
202
FAJ1
203
FAJ1
200
395
367
Ace
tyl
Ace
tyl t
rans
fera
setr
ansf
eras
e
Ace
tyl
Ace
tyl t
rans
fera
setr
ansf
eras
e
met
hyl
met
hyl t
rans
fera
setr
ansf
eras
e
met
hyl
met
hyl t
rans
fera
setr
ansf
eras
e
lpe3
lpeA
Fuco
seFu
cose
synt
hesis
synt
hesis
ABC
tra
nspo
rter
ABC
tra
nspo
rter
5
OO CH 3
O CH 3O H
H 3 C
O
CH 3
O
5 O-Acetyl groups
O HH OH O
O
OOO
H O
O HCO
O CH 3
O
O
H 3 C
O CH 3
H 3 C O
O
O H
O HO H
OO H
H 3 C
OH O
N H Ac
H O CH 3
OO
O
H OCH 2 O H
CO O H
H O
O
Core Lipid A
CH3
CH3
CH3
Kdo
QuiNAcManFuc
Fuc
dTal
GlcA
Fuc
25% 10%
367367
395395FAJ1201
FAJ1202
FAJ1203
FAJ1200
LipopolysaccharidesLipopolysaccharides: Structure, Synthesis, : Structure, Synthesis, FunctionFunction
Raetz, and Whitfield. 2002. Raetz, and Whitfield. 2002. LipopolysaccharideLipopolysaccharide EndotoxinsEndotoxins. Ann. Rev. . Ann. Rev. BiochemBiochem. 71:635. 71:635--700.700.