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Taxonomic group: bacteria / Proteobacteria (Phylum: Proteobacteria)
Host organism: Homo sapiens
Associated disease: acute otitis media [ICD11: AB00

, Life stage: adult]; chronic respiratory tract infections [ICD11: CA45

, ICD11: CB7Z

]; infection due to Haemophilus influenzae [ICD11: XN1P6

]

The structure was elucidated in this paper
NCBI PubMed ID: 21683945
Publication DOI: 10.1016/j.carres.2011.05.020
Journal NLM ID: 0043535
Publisher: Elsevier
Correspondence: E.K.H. Schweda <Elke.Schweda

ki.se>
Institutions: Karolinska Institutet, Clinical Research Centre, Huddinge, Sweden

The structure of lipopolysaccharide (LPS) expressed by non-typeable Haemophilus influenzae (NTHi) strains 1008 and 1247 has been investigated by mass spectrometry and NMR analyses on O-deacylated LPS and core oligosaccharide material. Both strains express the conserved triheptosyl inner core, [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] with PCho→6)-β-D-Glcp (GlcI) substituting the proximal heptose (HepI) at O-4. Strain 1247 expresses the common structural motifs of H. influenzae; globotetraose [β-D-GalpNAc-(1→3)-α-D-Galp-(1→4)-β-D-Galp-(1→4)-β-D-Glcp-(1→] and its truncated versions globoside [α-D-Galp-(1→4)-β-D-Galp-(1→4)-β-D-Glcp-(1→] and lactose [β-D-Galp-(1→4)-β-D-Glcp-(1→] linked to the terminal heptose of the inner core and GlcI. A genetically distinct NTHi strain, 1008, expresses identical structures to strain 1247 with the exception that it lacks GalNAc. A lpsA mutant of strain 1247 expressed LPS of reduced complexity that facilitated unambiguous structural determination of the oligosaccharide from HepI. By CE-ESI-MS/MS we identified disialylated glycoforms indicating disialyllactose [α-Neu5Ac-(2→8)-α-Neu5Ac-(2→3)-β-D-Gal-(1→4)-β-D-Glcp-(1→] as an extension from GlcI which is a novel finding for NTHi LPS.

Lipopolysaccharide, Haemophilus influenzae, otitis media, Digalactoside, disialyllactoside

Structure type: oligomer
Location inside paper: p.1892, table 6, HexNAcHex7, p.1894, fig.6
Aglycon: lipid A
Trivial name: HexNAcHex7 glycoform
Compound class: LPS
Contained glycoepitopes: IEDB_115009,IEDB_116046,IEDB_120354,IEDB_123890,IEDB_130648,IEDB_130650,IEDB_130651,IEDB_136044,IEDB_136906,IEDB_137338,IEDB_137472,IEDB_137473,IEDB_137777,IEDB_137779,IEDB_138949,IEDB_1391964,IEDB_140087,IEDB_140088,IEDB_140090,IEDB_140624,IEDB_141794,IEDB_142487,IEDB_142488,IEDB_144987,IEDB_146664,IEDB_151528,IEDB_152217,IEDB_190606,IEDB_2189047,IEDB_241118,IEDB_423106,IEDB_742247,IEDB_983931,SB_165,SB_166,SB_167,SB_178,SB_187,SB_192,SB_195,SB_21,SB_31,SB_6,SB_62,SB_7,SB_88

Methods: 13C NMR, 1H NMR, methylation, NMR-2D, GC-MS, HF solvolysis, ESI-MS, mild acid hydrolysis, de-O-acylation with hydrazine, composition analysis, NMR-1D, alkaline deacylation, HPAEC-PAD, CE-ESI-MS/MS, CE-ESI-MS, LC-ESI-MS
Comments, role: HexNAcHex7 glycoform core oligosaccharide from H. influenzae (NTHi) 1247. Published NMR assignment of terminal bDGalNAc C5 (67.2) is erroneous in Table 6.

Related record ID(s): 26373, 26977, 26994, 26995
NCBI Taxonomy refs (TaxIDs): 727
Show glycosyltransferases

NMR conditions: in D2O at 295 K [as TSV]

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6	C7
5,3,2,2,4,4	aDGalp	100.6	68.8	69.4	69.2	?	?
5,3,2,2,4	bDGalp	103.2	71.1	?	75.7	76.0	?
5,3,2,2	bDGlcp	102.2	72.6	74.4	78.7	71.9	60.3
5,3,2	aXLDmanHepp	99.8	78.4	?	?	?	?	?
5,3,6,0	xXEtN	62.8	40.2
5,3,6	P
5,3	aXLDmanHepp	99.1	79.1	?	?	?	74.82	62.7
5,4,4,4,4,3,2	Ac	?	22.6
5,4,4,4,4,3	bDGalpN	103.5	52.8	?	68.0	?	?
5,4,4,4,4	aDGalp	100.6	67.9	79.0	69.2	?	?
5,4,4,4	bDGalp	103.5	71.1	71.1	77.5	75.8	?
5,4,4	bDGlcp	103.5	73.9	74.34	79.484	74.34	60.7
5,4,6,0	xXCho	60.1	66.5	?
5,4,6	P
5,4	bDGlcp	103.3	73.2	72.4	79.4	71.8	64.5
5	aXLDmanHepp	97.0	?	?	74.4	?	68.0	?
4,0,0	xXEtN
4,0	%xXP?
4	P
	aXKdop


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6	H7
5,3,2,2,4,4	aDGalp	4.95	3.84	3.92	4.04	?	?
5,3,2,2,4	bDGalp	4.45	3.56	3.67	3.93	3.74	?
5,3,2,2	bDGlcp	4.68	3.34	3.71	3.69	3.66	3.83-4.00
5,3,2	aXLDmanHepp	5.09	4.20	3.94	?	?	?	?
5,3,6,0	xXEtN	4.12	3.22
5,3,6	P
5,3	aXLDmanHepp	5.58	4.25	3.96	?	3.71	4.55	3.89
5,4,4,4,4,3,2	Ac	-	2.05
5,4,4,4,4,3	bDGalpN	4.62	3.95	3.75	3.94	3.75	?
5,4,4,4,4	aDGalp	4.92	3.89	3.95	4.25	?	?
5,4,4,4	bDGalp	4.52	3.59	3.75	4.04	3.81	?
5,4,4	bDGlcp	4.50	3.38	3.75	3.75	3.75	4.01
5,4,6,0	xXCho	4.38	3.70	?
5,4,6	P
5,4	bDGlcp	4.54	3.43	3.72	3.66	3.63	4.19-4.34
5	aXLDmanHepp	5.03	3.99	?	4.20	?	4.19	?
4,0,0	xXEtN
4,0	%xXP?
4	P
	aXKdop


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6	C7/H7
5,3,2,2,4,4	aDGalp	100.6/4.95	68.8/3.84	69.4/3.92	69.2/4.04	?/?	?/?
5,3,2,2,4	bDGalp	103.2/4.45	71.1/3.56	?/3.67	75.7/3.93	76.0/3.74	?/?
5,3,2,2	bDGlcp	102.2/4.68	72.6/3.34	74.4/3.71	78.7/3.69	71.9/3.66	60.3/3.83-4.00
5,3,2	aXLDmanHepp	99.8/5.09	78.4/4.20	?/3.94	?/?	?/?	?/?	?/?
5,3,6,0	xXEtN	62.8/4.12	40.2/3.22
5,3,6	P
5,3	aXLDmanHepp	99.1/5.58	79.1/4.25	?/3.96	?/?	?/3.71	74.82/4.55	62.7/3.89
5,4,4,4,4,3,2	Ac		22.6/2.05
5,4,4,4,4,3	bDGalpN	103.5/4.62	52.8/3.95	?/3.75	68.0/3.94	?/3.75	?/?
5,4,4,4,4	aDGalp	100.6/4.92	67.9/3.89	79.0/3.95	69.2/4.25	?/?	?/?
5,4,4,4	bDGalp	103.5/4.52	71.1/3.59	71.1/3.75	77.5/4.04	75.8/3.81	?/?
5,4,4	bDGlcp	103.5/4.50	73.9/3.38	74.34/3.75	79.484/3.75	74.34/3.75	60.7/4.01
5,4,6,0	xXCho	60.1/4.38	66.5/3.70	?/?
5,4,6	P
5,4	bDGlcp	103.3/4.54	73.2/3.43	72.4/3.72	79.4/3.66	71.8/3.63	64.5/4.19-4.34
5	aXLDmanHepp	97.0/5.03	?/3.99	?/?	74.4/4.20	?/?	68.0/4.19	?/?
4,0,0	xXEtN
4,0	%xXP?
4	P
	aXKdop

¹H NMR data:

Linkage	Residue	H1	H2	H3	H4	H5	H6	H7
5,3,2,2,4,4	aDGalp	4.95	3.84	3.92	4.04	?	?
5,3,2,2,4	bDGalp	4.45	3.56	3.67	3.93	3.74	?
5,3,2,2	bDGlcp	4.68	3.34	3.71	3.69	3.66	3.83 4.00
5,3,2	aXLDmanHepp	5.09	4.20	3.94	?	?	?	?
5,3,6,0	xXEtN	4.12	3.22
5,3,6	P
5,3	aXLDmanHepp	5.58	4.25	3.96	?	3.71	4.55	3.89
5,4,4,4,4,3,2	Ac		2.05
5,4,4,4,4,3	bDGalpN	4.62	3.95	3.75	3.94	3.75	?
5,4,4,4,4	aDGalp	4.92	3.89	3.95	4.25	?	?
5,4,4,4	bDGalp	4.52	3.59	3.75	4.04	3.81	?
5,4,4	bDGlcp	4.50	3.38	3.75	3.75	3.75	4.01
5,4,6,0	xXCho	4.38	3.70	?
5,4,6	P
5,4	bDGlcp	4.54	3.43	3.72	3.66	3.63	4.19 4.34
5	aXLDmanHepp	5.03	3.99	?	4.20	?	4.19	?
4,0,0	xXEtN
4,0	%xXP?
4	P
	aXKdop

¹³C NMR data:

Linkage	Residue	C1	C2	C3	C4	C5	C6	C7
5,3,2,2,4,4	aDGalp	100.6	68.8	69.4	69.2	?	?
5,3,2,2,4	bDGalp	103.2	71.1	?	75.7	76.0	?
5,3,2,2	bDGlcp	102.2	72.6	74.4	78.7	71.9	60.3
5,3,2	aXLDmanHepp	99.8	78.4	?	?	?	?	?
5,3,6,0	xXEtN	62.8	40.2
5,3,6	P
5,3	aXLDmanHepp	99.1	79.1	?	?	?	74.82	62.7
5,4,4,4,4,3,2	Ac	?	22.6
5,4,4,4,4,3	bDGalpN	103.5	52.8	?	68.0	?	?
5,4,4,4,4	aDGalp	100.6	67.9	79.0	69.2	?	?
5,4,4,4	bDGalp	103.5	71.1	71.1	77.5	75.8	?
5,4,4	bDGlcp	103.5	73.9	74.34	79.484	74.34	60.7
5,4,6,0	xXCho	60.1	66.5	?
5,4,6	P
5,4	bDGlcp	103.3	73.2	72.4	79.4	71.8	64.5
5	aXLDmanHepp	97.0	?	?	74.4	?	68.0	?
4,0,0	xXEtN
4,0	%xXP?
4	P
	aXKdop

The spectrum also has 24 signals at unknown positions (not plotted).

There is only one chemically distinct structure:

CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@H](O[C@H]3[C@@H](CO)O[C@@H](O[C@@H]4[C@@H](CO)O[C@@H](O[C@@H]5[C@@H](COP(=O)(O)OCC[N+](C)(C)C)O[C@@H](O[C@@H]6[C@@H]([C@@H](O)CO)O[C@H](O[C@@H]7[C@H](OP(=O)(O)OP(=O)(O)OCCN)C[C@](O)(C(=O)O)O[C@@H]7[C@H](O)CO)[C@@H](O)[C@H]6O[C@H]6O[C@H]([C@H](CO)OP(=O)(O)OCCN)[C@@H](O)[C@H](O)[C@@H]6O[C@H]6O[C@H]([C@@H](O)CO)[C@@H](O)[C@H](O)[C@@H]6O[C@@H]6O[C@H](CO)[C@@H](O[C@@H]7O[C@H](CO)[C@H](O[C@H]8O[C@H](CO)[C@H](O)[C@H](O)[C@H]8O)[C@H](O)[C@H]7O)[C@H](O)[C@H]6O)[C@H](O)[C@H]5O)[C@H](O)[C@H]4O)[C@H](O)[C@H]3O)[C@@H]2O)O[C@H](CO)[C@H](O)[C@@H]1O

SVG MW SMILES 3D mol Ionize