Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Host organism: Homo sapiens
Organ / tissue: middle earAssociated disease: acute otitis media [ICD11:
AB00 
, Life stage: adult];
infection due to Haemophilus influenzae [ICD11:
XN1P6 ]
The structure was elucidated in this paperNCBI PubMed ID: 18465844Publication DOI: 10.1021/bi702510bJournal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: Susanna.Lundstrom
ki.se
Institutions: Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada, K1A 0R6, Clinical Research Centre, Karolinska Institutet and University College of South Stockholm, Novum, Huddinge, Sweden, Molecular Infectious Diseases Group, University of Oxford, Department of Paediatrics, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, U.K.
We here report the lipopolysaccharide (LPS) structures expressed by nontypeable Haemophilus influenzae R2846, a strain whose complete genome sequence has recently been obtained. Results were obtained by using NMR techniques and ESI-MS on O-deacylated LPS and core oligosaccharide material (OS) as well as ESI-MS (n) on permethylated dephosphorylated OS. A β-D-Glcp-(1→4)-D-α-D-Hepp-(1→6)-β-D-Glcp-(1→4) unit was found linked to the proximal heptose (HepI) of the conserved triheptosyl inner-core moiety, l-α-D-Hepp-(1→2)-[PEtn→6]-L-α-D-Hepp-(1→3)-L-α-D-Hepp-(1→5)-[PPEtn→4]-α-Kdo-(2→6)-lipid A. The β-D-Glcp (GlcI) linked to HepI was also branched with oligosaccharide extensions from O-4 and O-6. O-4 of GlcI was substituted with sialyllacto- N-neotetraose [α-Neu5Ac-(2→3)-β-D-Galp-(1→4)-β-D-GlcpNAc-(1→3)-β-D-Galp-(1→4)-β-D-Glcp-(1→] and the related structure [(PEtn→6)-α-D-GalpNAc-(1→6)-β-D-Galp-(1→4)-β-D-GlcpNAc-(1→3)-β-D-Galp-(1→4)-β-D-Glcp-(1→]. The distal heptose (HepIII) was substituted at O-2 by β-D-Gal. Phosphate, phosphoethanolamine, phosphocholine, acetate, and glycine were found to substitute the core oligosaccharide. Two heptosyltransferase genes, losB1 and losB2, have been identified from the R2846 genome sequence and are candidates to add the noncore heptose to the LPS. Mutant strain R2846 losB1 did not show dd-heptose in the extension from HepI but still contained minor quantities of ld-heptose at the same position, indicating that the losB1 gene is required to add dd-heptose to GlcI. The LPS from strain R2846 losB1/ losB2 expressed no noncore heptose, consistent with losB2 directing the addition of LD-heptose.
Lipopolysaccharide, NMR, Haemophilus influenzae, core oligosaccharide, structural analysis
Structure type: oligomer
Location inside paper: p.6031, chart 1
Trivial name: glycoform Hex3Hep4
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_136044,IEDB_137472,IEDB_137779,IEDB_139428,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_141794,IEDB_142488,IEDB_146664,IEDB_190606,IEDB_2189046,IEDB_2189047,IEDB_983931,SB_165,SB_166,SB_187,SB_192,SB_195,SB_7,SB_88
Methods: 13C NMR, 1H NMR, GLC-MS, NMR-2D, HF solvolysis, sugar analysis, 31P NMR, ESI-MS, mild acid hydrolysis, ESI-MS/MS, NMR-1D, genetic methods, HPLC, alkaline hydrolysis, CE-ESI-MS
Biosynthesis and genetic data: genetic data
Comments, role: O-deacylated oligosaccharide OS3.
Related record ID(s): 23118, 23119, 23120, 23121, 23122, 23123, 31135, 31136
NCBI Taxonomy refs (TaxIDs): 262727
Show glycosyltransferases
NMR conditions: in D2O at 298 K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6 C7
5,3,2,2 bDGalp 103.8 70.8 72.8 69.2 75.7 ?
5,3,2 aXLDmanHepp 100.1 79.7 70.0 ? ? ? ?
5,3,6,0 xXEtN
5,3,6 P
5,3 aXLDmanHepp 99.2 79.6 69.3 ? 71.6 75.3 62.4
5,4,6,4 bDGlcp 103.2 73.8 76.0 70.2 76.9 61.3
5,4,6 aXDDmanHepp 99.3 69.9 70.5 78.6 71.0 ? ?
5,4 bDGlcp 103.6 74.1 77.2 70.6 74.1 66.0
5 aXLDmanHepp 97.1 70.9 72.8 73.8 ? 68.2 ?
x?Sug?
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6 H7
5,3,2,2 bDGalp 4.40 3.59 3.70 3.92 3.75 ?
5,3,2 aXLDmanHepp 4.99 4.14 4.00 ? ? ? ?
5,3,6,0 xXEtN
5,3,6 P
5,3 aXLDmanHepp 5.68 4.18 3.93 ? 3.75 4.57 3.72-3.90
5,4,6,4 bDGlcp 4.56 3.31 3.55 3.41 3.55 3.74-3.96
5,4,6 aXDDmanHepp 4.96 4.11 3.93 3.94 4.18 ? ?
5,4 bDGlcp 4.50 3.40 3.44 3.54 3.59 3.85-4.03
5 aXLDmanHepp 5.04-5.15 4.00-4.06 4.01 ? 4.11 ? ?
x?Sug?
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6 C7/H7
5,3,2,2 bDGalp 103.8/4.40 70.8/3.59 72.8/3.70 69.2/3.92 75.7/3.75 ?/?
5,3,2 aXLDmanHepp 100.1/4.99 79.7/4.14 70.0/4.00 ?/? ?/? ?/? ?/?
5,3,6,0 xXEtN
5,3,6 P
5,3 aXLDmanHepp 99.2/5.68 79.6/4.18 69.3/3.93 ?/? 71.6/3.75 75.3/4.57 62.4/3.72-3.90
5,4,6,4 bDGlcp 103.2/4.56 73.8/3.31 76.0/3.55 70.2/3.41 76.9/3.55 61.3/3.74-3.96
5,4,6 aXDDmanHepp 99.3/4.96 69.9/4.11 70.5/3.93 78.6/3.94 71.0/4.18 ?/? ?/?
5,4 bDGlcp 103.6/4.50 74.1/3.40 77.2/3.44 70.6/3.54 74.1/3.59 66.0/3.85-4.03
5 aXLDmanHepp 97.1/5.04-5.15 70.9/4.00-4.06 72.8/4.01 73.8/? ?/4.11 68.2/? ?/?
x?Sug?
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 | H7 |
|---|
| 5,3,2,2 | bDGalp | 4.40 | 3.59 | 3.70 | 3.92 | 3.75 | ? | |
| 5,3,2 | aXLDmanHepp | 4.99 | 4.14 | 4.00 | ? | ? | ? | ? |
| 5,3,6,0 | xXEtN | |
| 5,3,6 | P | |
| 5,3 | aXLDmanHepp | 5.68 | 4.18 | 3.93 | ? | 3.75 | 4.57 | 3.72
3.90 |
| 5,4,6,4 | bDGlcp | 4.56 | 3.31 | 3.55 | 3.41 | 3.55 | 3.74
3.96 | |
| 5,4,6 | aXDDmanHepp | 4.96 | 4.11 | 3.93 | 3.94 | 4.18 | ? | ? |
| 5,4 | bDGlcp | 4.50 | 3.40 | 3.44 | 3.54 | 3.59 | 3.85
4.03 | |
| 5 | aXLDmanHepp | 5.04
5.15 | 4.00
4.06 | 4.01 | ? | 4.11 | ? | ? |
| | x?Sug? | |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 | C7 |
|---|
| 5,3,2,2 | bDGalp | 103.8 | 70.8 | 72.8 | 69.2 | 75.7 | ? | |
| 5,3,2 | aXLDmanHepp | 100.1 | 79.7 | 70.0 | ? | ? | ? | ? |
| 5,3,6,0 | xXEtN | |
| 5,3,6 | P | |
| 5,3 | aXLDmanHepp | 99.2 | 79.6 | 69.3 | ? | 71.6 | 75.3 | 62.4 |
| 5,4,6,4 | bDGlcp | 103.2 | 73.8 | 76.0 | 70.2 | 76.9 | 61.3 | |
| 5,4,6 | aXDDmanHepp | 99.3 | 69.9 | 70.5 | 78.6 | 71.0 | ? | ? |
| 5,4 | bDGlcp | 103.6 | 74.1 | 77.2 | 70.6 | 74.1 | 66.0 | |
| 5 | aXLDmanHepp | 97.1 | 70.9 | 72.8 | 73.8 | ? | 68.2 | ? |
| | x?Sug? | |
|
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The spectrum also has 10 signals at unknown positions (not plotted). |
There is only one chemically distinct structure:
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Found 1 record.
Displayed record 1
