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Greenfield LK, Richards MR, Li J, Wakarchuk WW, Lowary TL, Whitfield C
Biosynthesis of the polymannose lipopolysaccharide O antigens from Escherichia coli serotypes O8 and O9a requires a unique combination of single- and multi-active site mannosyltransferases
Journal of Biological Chemistry 287(42) (2012)
35078-35091
Geobacillus stearothermophilus NRS 2004/3a
(Ancestor NCBI TaxID 1422,
species name lookup)
Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
NCBI PubMed ID: 22875852Publication DOI: 10.1074/jbc.M112.401000DAJournal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: cwhitfie
uoguelph.ca
Institutions: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada
The Escherichia coli O9a and O8 O-antigen serotypes represent model systems for the ABC transporter-dependent synthesis of bacterial polysaccharides. The O9a and O8 antigens are linear mannose homopolymers containing conserved reducing termini (the primer-adaptor), a serotype-specific repeat-unit domain, and a terminator. Synthesis of these glycans occurs on the polyisoprenoid lipid-linked primer, undecaprenyl pyrophosphoryl-GlcpNAc, by two conserved mannosyltransferases, WbdC and WbdB, and a serotype-specific mannosyltransferase, WbdA. The glycan structure and pattern of conservation in the O9a and O8 mannosyltransferases are not consistent with the existing model of O9a biosynthesis. Here we establish a revised pathway using a combination of in vivo (mutant complementation) experiments and in vitro strategies with purified enzymes and synthetic acceptors. WbdC and WbdB synthesize the adaptor region, where they transfer one and two α-(1→3)-linked mannose residues, respectively. The WbdA enzymes are solely responsible for forming the repeat-unit domains of these O antigens. WbdA(O9a) has two predicted active sites and polymerizes a tetrasaccharide repeat unit containing two α-(1→3)- and two α-(1→2)-linked mannopyranose residues. In contrast, WbdA(O8) polymerizes trisaccharide repeat units containing single α-(1→3), α-(1→2) and β-(1→2)-mannopyranoses. These studies illustrate assembly systems exploiting several mannosyltransferases with flexible active sites, arranged in single and multi-domain formats
biosynthesis, Escherichia coli, O-antigens, mannosyltransferases
Structure type: polymer chemical repeating unit
Location inside paper: p.35089
Trivial name: S-layer glycoprotein, S-layer glycan
Compound class: surface layer glycoprotein
Contained glycoepitopes: IEDB_133754,IEDB_136105,IEDB_225177,IEDB_885823
Methods: 13C NMR, 1H NMR, NMR-2D, SDS-PAGE, DNA techniques, MALDI-TOF MS, NMR-1D, biochemical methods, CE, computer analysis with CASPER
Synthetic data: enzymatic
Related record ID(s): 27164, 28347, 28348, 28349
NCBI Taxonomy refs (TaxIDs): 1422Reference(s) to other database(s): GTC:G04332YD, GlycomeDB:
5425
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Christian R, Schulz G, Unger FM, Messner P, Küpcü Z, Sleytr UB
Structure of a rhamnan from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a
Carbohydrate Research 150 (1986)
265-272
Bacillus stearothermophilus
(later renamed to: Geobacillus stearothermophilus)
(NCBI TaxID 1422,
species name lookup)
Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
Organ / tissue: cell surface
Journal NLM ID: 0043535Publisher: Elsevier
Structure type: polymer chemical repeating unit
Trivial name: S-layer glycoprotein, S-layer glycan
Compound class: surface layer glycoprotein
Contained glycoepitopes: IEDB_133754,IEDB_136105,IEDB_225177,IEDB_885823
Comments, role: published polymerization frame was shifted for conformity with other records
Related record ID(s): 115132
NCBI Taxonomy refs (TaxIDs): 1422Reference(s) to other database(s): GTC:G04332YD, GlycomeDB:
5425
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Messner P, Sleytr UB, Christian R, Schulz G, Unger FM
Isolation and structure determination of a diacetamidodideoxyuronic acid containing glycan chain from the S-layer glycoprotein of Bacillus stearothermophilus NRS 2004/3a
Carbohydrate Research 168 (1987)
211-218
Bacillus stearothermophilus NRS 2004/3a
(later renamed to: Geobacillus stearothermophilus NRS 2004/3a)
(Ancestor NCBI TaxID 1422,
species name lookup)
Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
Organ / tissue: cell surface
The structure was elucidated in this paperNCBI PubMed ID: 3427581Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Ultrastrukturforschung, Universität für Bodenkultur, A-1180 Wien Austria
A glycan isolated from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a was shown by 1H- and 13C-n.m.r. spectroscopy to have the tetrasaccharide repeating-unit →4)-β-ManpA2,3(NAc)2-(1→3)-α-GlcpNAc-(1→4)-β- ManpA2,3(NAc)-(1→6)-α-Glcp(1→.
Structure type: polymer chemical repeating unit
Location inside paper: GP-I
Trivial name: S-layer glycoprotein, S-layer glycan
Compound class: surface layer glycoprotein
Contained glycoepitopes: IEDB_133754,IEDB_136105,IEDB_225177,IEDB_885823
Methods: 13C NMR, 1H NMR
Comments, role: published polymerization frame was shifted for conformity with other records.
Related record ID(s): 114763, 115133
NCBI Taxonomy refs (TaxIDs): 1422Reference(s) to other database(s): GTC:G04332YD, GlycomeDB:
5425
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There is only one chemically distinct structure:
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Messner P, Sleytr UB, Christian R, Schulz G, Unger FM
Isolation and structure determination of a diacetamidodideoxyuronic acid containing glycan chain from the S-layer glycoprotein of Bacillus stearothermophilus NRS 2004/3a
Carbohydrate Research 168 (1987)
211-218
|
-4)-b-D-ManpNAc3NAcA-(1-3)-a-D-GlcpNAc-(1-4)-b-D-ManpNAc3NAcA-(1-6)-a-D-Glcp-(1- |
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Bacillus stearothermophilus
(later renamed to: Geobacillus stearothermophilus)
(NCBI TaxID 1422,
species name lookup)
Taxonomic group: bacteria / Firmicutes
(Phylum: Firmicutes)
Organ / tissue: cell surface
The structure was elucidated in this paperNCBI PubMed ID: 3427581Journal NLM ID: 0043535Publisher: Elsevier
Institutions: Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Ultrastrukturforschung, Universität für Bodenkultur, A-1180 Wien Austria
A glycan isolated from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a was shown by 1H- and 13C-n.m.r. spectroscopy to have the tetrasaccharide repeating-unit →4)-β-ManpA2,3(NAc)2-(1→3)-α-GlcpNAc-(1→4)-β- ManpA2,3(NAc)-(1→6)-α-Glcp(1→.
Structure type: polymer chemical repeating unit
Location inside paper: GP-II
Trivial name: S-layer glycoprotein
Compound class: surface layer glycoprotein
Contained glycoepitopes: IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151531,IEDB_983931,SB_192
Methods: 13C NMR, 1H NMR
Related record ID(s): 115132
NCBI Taxonomy refs (TaxIDs): 1422Reference(s) to other database(s): GTC:G09633JJ, GlycomeDB:
5589
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There is only one chemically distinct structure:
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