Journal
CARBOHYDRATE RESEARCH
Volume 498, Issue -, Pages -Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carres.2020.108051
Keywords
Escherichia coli; Lipopolysaccharide; NMR spectroscopy; Shigella boydii
Funding
- Swedish Research Council [2017-03703]
- Knut and Alice Wallenberg Foundation
- Vinnova [2017-03703] Funding Source: Vinnova
- Swedish Research Council [2017-03703] Funding Source: Swedish Research Council
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The structure of the O-antigen from Escherichia coli reference strain O188 (E. coli O188:H10) has been investigated. The lipopolysaccharide shows a typical nonrandom modal chain-length distribution and the sugar and absolute configuration analysis revealed D-Man, D-Glc, D-GlcN and D-GlcA as major components. The structure of the O-specific polysaccharide was determined using one- and two-dimensional H-1 and C-13 NMR spectroscopy experiments, where inter-residue correlations were identified by H-1,C-13-heteronuclear multiple-bond correlation and H-1,H-1-NOESY experiments, which revealed that it consists of pentasaccharide repeating units with the -> 4)-beta-D-GlcpA-(1 -> 2)-beta-D-Manp-(1 -> 4)-beta-D-Manp-(1 -> 3)-beta-D-GlcpNAc-(1 -> following structure: vertical bar alpha-D-Galp-(1 -> 3) Biosynthetic aspects and NMR analysis are consistent with the presented structure as the biological repeating unit. The O-antigen of Shigella boydii type 16 differs only in that it carries O-acetyl groups to similar to 50% at O6 of the branchpoint mannose residues. A molecular model of the E. coli O188 O-antigen containing 20 repeating units extends similar to 100 angstrom, which is similar to the height of the periplasmic portion of polysaccharide co-polymerase Wzz proteins that regulate the O-antigen chain length of lipopolysaccharides in the Wzx/Wzy biosynthetic pathway.
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