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Taxonomic group: bacteria / Proteobacteria (Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11: XN6P4

]

The structure was elucidated in this paper
NCBI PubMed ID: 32839412
Publication DOI: 10.3390/ijms21176038
Journal NLM ID: 101092791
Publisher: Basel, Switzerland: MDPI
Correspondence: jolanta.lukasiewicz

hirszfeld.pl
Institutions: Laboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland

Enterobacterial common antigen (ECA) is a conserved surface antigen characteristic for Enterobacteriaceae. It is consisting of trisaccharide repeating unit, →3)-α-d-Fucp4NAc-(1→4)-β-d-ManpNAcA-(1→4)-α-d-GlcpNAc-(1→, where prevailing forms include ECA linked to phosphatidylglycerol (ECAPG) and cyclic ECA (ECACYC). Lipopolysaccharide (LPS)-associated form (ECALPS) has been proved to date only for rough Shigella sonnei phase II. Depending on the structure organization, ECA constitutes surface antigen (ECAPG and ECALPS) or maintains the outer membrane permeability barrier (ECACYC). The existence of LPS was hypothesized in the 1960-80s on the basis of serological observations. Only a few Escherichia coli strains (i.e., R1, R2, R3, R4, and K-12) have led to the generation of anti-ECA antibodies upon immunization, excluding ECAPG as an immunogen and conjecturing ECALPS as the only immunogenic form. Here, we presented a structural survey of ECALPS in E. coli R1, R2, R3, and R4 to correlate previous serological observations with the presence of ECALPS. The low yields of ECALPS were identified in the R1, R2, and R4 strains, where ECA occupied outer core residues of LPS that used to be substituted by O-specific polysaccharide in the case of smooth LPS. Previously published observations and hypotheses regarding the immunogenicity and biosynthesis of ECALPS were discussed and correlated with presented herein structural data.

Lipopolysaccharide, NMR, LPS, enterobacterial common antigen, ECA, endotoxin, mass spectrometry, ECALPS

Structure type: oligomer
Location inside paper: Fig.3a, table 2, E. coli R1
Trivial name: ECA(LPS)
Compound class: ECA-core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130670,IEDB_131186,IEDB_133751,IEDB_135813,IEDB_135818,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_140088,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_153543,IEDB_190606,IEDB_2189047,IEDB_226811,IEDB_983931,SB_192,SB_7

Methods: 13C NMR, 1H NMR, NMR-2D, ESI-MS, MS/MS, MALDI-TOF MS
Comments, role: ECA-core glycoform in fractions 5 of E. coli R1.

Related record ID(s): 5138, 5139, 5140, 5141, 32158
NCBI Taxonomy refs (TaxIDs): 562
Reference(s) to other database(s): GTC:G32203GI
Show glycosyltransferases

NMR conditions: in D2O at 303 K [as TSV]

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6	C7	C8
5,3,3,3,2,2	aDGalp	96.6	69.2	70.2	70.2	72.1	62.1
5,3,3,3,2	aDGalp	92.1	73.2	68.9	70.9	72.2	62.1
5,3,3,3,3,3,4,4,4	Ac
5,3,3,3,3,3,4,4	aDFucp4N	99.7	69.3	69.2	54.7	66.7	16.3
5,3,3,3,3,3,4,2	Ac
5,3,3,3,3,3,4	bDManpNA	99.8	54.3	73.3	74.9	77.2	175.1
5,3,3,3,3,3,2	Ac
5,3,3,3,3,3	bDGlcpN	102.4	56.4	72.8	79.6	75.2	61.0
5,3,3,3,3	bDGlcp	103.2	73.7	85.5	69.0	76.3	61.5
5,3,3,3	aDGlcp	95.4	73.5	78.7	68.8	72.1	61.1
5,3,3	aDGlcp	102.1	71.2	76.8	71.3	73.2	60.7
5,3,4	P
5,3,7	aXLDmanHepp	100.3	70.9	71.5	67.0	72.0	69.6	63.7
5,3	aXLDmanHepp	103.7	70.6	79.9	69.5	73.3	68.8	68.5
5,4,0,0	xXEtN	63.2	40.8
5,4,0	P
5,4	P
5	aXLDmanHepp	100.2	71.8	78.6	72.4	72.1	69.4	63.9
	aXKdop	?	96.2	34.2	66.4	73.4	69.9	73.0	64.9


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6	H7	H8
5,3,3,3,2,2	aDGalp	5.32	3.86	3.96	4.00	4.14	3.75
5,3,3,3,2	aDGalp	5.63	4.00	4.20	3.99	4.15	3.76
5,3,3,3,3,3,4,4,4	Ac
5,3,3,3,3,3,4,4	aDFucp4N	5.36	3.65	3.97	4.21	4.19	1.06
5,3,3,3,3,3,4,2	Ac
5,3,3,3,3,3,4	bDManpNA	4.94	4.51	4.08	3.84	3.88	-
5,3,3,3,3,3,2	Ac
5,3,3,3,3,3	bDGlcpN	4.79	3.76	3.75	3.70	3.55	3.73-3.86
5,3,3,3,3	bDGlcp	4.74	3.39	3.70	3.50	3.45	3.75-3.91
5,3,3,3	aDGlcp	5.81	3.88	4.18	3.58	4.11	3.79-4.00
5,3,3	aDGlcp	5.21	3.68	4.09	3.80	3.92	3.81-3.92
5,3,4	P
5,3,7	aXLDmanHepp	5.00	3.94	3.89	3.86	3.63	4.05	3.64-3.75
5,3	aXLDmanHepp	5.11	4.40	4.12	4.42	3.81	4.23	3.61-3.77
5,4,0,0	xXEtN	4.21	3.30
5,4,0	P
5,4	P
5	aXLDmanHepp	5.21	4.02	4.09	4.64	4.24	4.11	3.73
	aXKdop	-	-	1.91-2.27	4.13	4.18	3.70	3.81	3.47-3.94


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6	C7/H7	C8/H8
5,3,3,3,2,2	aDGalp	96.6/5.32	69.2/3.86	70.2/3.96	70.2/4.00	72.1/4.14	62.1/3.75
5,3,3,3,2	aDGalp	92.1/5.63	73.2/4.00	68.9/4.20	70.9/3.99	72.2/4.15	62.1/3.76
5,3,3,3,3,3,4,4,4	Ac
5,3,3,3,3,3,4,4	aDFucp4N	99.7/5.36	69.3/3.65	69.2/3.97	54.7/4.21	66.7/4.19	16.3/1.06
5,3,3,3,3,3,4,2	Ac
5,3,3,3,3,3,4	bDManpNA	99.8/4.94	54.3/4.51	73.3/4.08	74.9/3.84	77.2/3.88	
5,3,3,3,3,3,2	Ac
5,3,3,3,3,3	bDGlcpN	102.4/4.79	56.4/3.76	72.8/3.75	79.6/3.70	75.2/3.55	61.0/3.73-3.86
5,3,3,3,3	bDGlcp	103.2/4.74	73.7/3.39	85.5/3.70	69.0/3.50	76.3/3.45	61.5/3.75-3.91
5,3,3,3	aDGlcp	95.4/5.81	73.5/3.88	78.7/4.18	68.8/3.58	72.1/4.11	61.1/3.79-4.00
5,3,3	aDGlcp	102.1/5.21	71.2/3.68	76.8/4.09	71.3/3.80	73.2/3.92	60.7/3.81-3.92
5,3,4	P
5,3,7	aXLDmanHepp	100.3/5.00	70.9/3.94	71.5/3.89	67.0/3.86	72.0/3.63	69.6/4.05	63.7/3.64-3.75
5,3	aXLDmanHepp	103.7/5.11	70.6/4.40	79.9/4.12	69.5/4.42	73.3/3.81	68.8/4.23	68.5/3.61-3.77
5,4,0,0	xXEtN	63.2/4.21	40.8/3.30
5,4,0	P
5,4	P
5	aXLDmanHepp	100.2/5.21	71.8/4.02	78.6/4.09	72.4/4.64	72.1/4.24	69.4/4.11	63.9/3.73
	aXKdop			34.2/1.91-2.27	66.4/4.13	73.4/4.18	69.9/3.70	73.0/3.81	64.9/3.47-3.94

¹H NMR data:

Linkage	Residue	H1	H2	H3	H4	H5	H6	H7	H8
5,3,3,3,2,2	aDGalp	5.32	3.86	3.96	4.00	4.14	3.75
5,3,3,3,2	aDGalp	5.63	4.00	4.20	3.99	4.15	3.76
5,3,3,3,3,3,4,4,4	Ac
5,3,3,3,3,3,4,4	aDFucp4N	5.36	3.65	3.97	4.21	4.19	1.06
5,3,3,3,3,3,4,2	Ac
5,3,3,3,3,3,4	bDManpNA	4.94	4.51	4.08	3.84	3.88
5,3,3,3,3,3,2	Ac
5,3,3,3,3,3	bDGlcpN	4.79	3.76	3.75	3.70	3.55	3.73 3.86
5,3,3,3,3	bDGlcp	4.74	3.39	3.70	3.50	3.45	3.75 3.91
5,3,3,3	aDGlcp	5.81	3.88	4.18	3.58	4.11	3.79 4.00
5,3,3	aDGlcp	5.21	3.68	4.09	3.80	3.92	3.81 3.92
5,3,4	P
5,3,7	aXLDmanHepp	5.00	3.94	3.89	3.86	3.63	4.05	3.64 3.75
5,3	aXLDmanHepp	5.11	4.40	4.12	4.42	3.81	4.23	3.61 3.77
5,4,0,0	xXEtN	4.21	3.30
5,4,0	P
5,4	P
5	aXLDmanHepp	5.21	4.02	4.09	4.64	4.24	4.11	3.73
	aXKdop			1.91 2.27	4.13	4.18	3.70	3.81	3.47 3.94

¹³C NMR data:

Linkage	Residue	C1	C2	C3	C4	C5	C6	C7	C8
5,3,3,3,2,2	aDGalp	96.6	69.2	70.2	70.2	72.1	62.1
5,3,3,3,2	aDGalp	92.1	73.2	68.9	70.9	72.2	62.1
5,3,3,3,3,3,4,4,4	Ac
5,3,3,3,3,3,4,4	aDFucp4N	99.7	69.3	69.2	54.7	66.7	16.3
5,3,3,3,3,3,4,2	Ac
5,3,3,3,3,3,4	bDManpNA	99.8	54.3	73.3	74.9	77.2	175.1
5,3,3,3,3,3,2	Ac
5,3,3,3,3,3	bDGlcpN	102.4	56.4	72.8	79.6	75.2	61.0
5,3,3,3,3	bDGlcp	103.2	73.7	85.5	69.0	76.3	61.5
5,3,3,3	aDGlcp	95.4	73.5	78.7	68.8	72.1	61.1
5,3,3	aDGlcp	102.1	71.2	76.8	71.3	73.2	60.7
5,3,4	P
5,3,7	aXLDmanHepp	100.3	70.9	71.5	67.0	72.0	69.6	63.7
5,3	aXLDmanHepp	103.7	70.6	79.9	69.5	73.3	68.8	68.5
5,4,0,0	xXEtN	63.2	40.8
5,4,0	P
5,4	P
5	aXLDmanHepp	100.2	71.8	78.6	72.4	72.1	69.4	63.9
	aXKdop	?	96.2	34.2	66.4	73.4	69.9	73.0	64.9

The spectrum also has 1 signal at unknown position (not plotted).

There is only one chemically distinct structure:

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

SVG MW SMILES 3D mol Ionize