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Gozdziewicz TK, Maciejewska A, Tsybulska A, Lugowski C, Lukasiewicz J
A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification
International Journal of Molecular Sciences 22(2) (2021)
701
| Cyclic
-3)-a-D-Fucp4NAc-(1-4)-b-D-ManpNAcA-(1-4)-a-D-GlcpNAc6(%)Ac-(1- |
Show graphically |
Shigella sonnei PCM 1985
(Ancestor NCBI TaxID 624,
species name lookup)
Escherichia coli R1 F470
(Ancestor NCBI TaxID 562,
species name lookup)
Escherichia coli K12 W3110
(NCBI TaxID 316407,
species name lookup)
Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Shigella sonnei [ICD11:
XN9M9 
];
infection due to Escherichia coli [ICD11:
XN6P4 ]
The structure was elucidated in this paperNCBI PubMed ID: 33445776Publication DOI: 10.3390/ijms22020701Journal NLM ID: 101092791Publisher: 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 antigen expressed by enterobacteria. It is built by trisaccharide repeating units: →3)-α-D-Fucp4NAc-(1→4)-β-D-ManpNAcA-(1→4)-α-D-GlcpNAc-(1→ and occurs in three forms: as surface-bound linear polysaccharides linked to a phosphoglyceride (ECAPG) or lipopolysaccharide - endotoxin (ECALPS), and cyclic form (ECACYC). ECA maintains, outer membrane integrity, immunogenicity, and viability of enterobacteria. A supernatant obtained after LPS ultracentrifugation was reported as a source for ECA isolation, but it has never been assessed for detailed composition besides ECACYC. We used mild acid hydrolysis and gel filtration, or zwitterionic-hydrophilic interaction liquid (ZIC®HILIC) chromatography combined with mass spectrometry for purification, fractionation, and structural analysis of rough Shigella sonnei and Escherichia coli R1 and K12 crude LPS preparations. Presented work is the first report concerning complex characteristic of all ECA forms present in LPS-derived supernatants. We demonstrated high heterogeneity of the supernatant-derived ECA that contaminate LPS purified by ultracentrifugation. Not only previously reported O-acetylated tetrameric, pentameric, and hexameric ECACYC have been identified, but also devoid of lipid moiety linear ECA built from 7 to 11 repeating units. Described results were common for all selected strains. The origin of linear ECA is discussed against the current knowledge about ECAPG and ECALPS.
Lipopolysaccharide, LPS, enterobacterial common antigen, ECA, mass spectrometry, cyclic ECA, ECAPG
Structure type: cyclic polymer repeating unit ; n=4-6
Location inside paper: abstract, Fig. 3, table 2, table 3
Trivial name: ECA(CYC) cyclic form
Compound class: Enterobacterial common antigen
Contained glycoepitopes: IEDB_141807,IEDB_151531
Methods: gel filtration, ESI-MS, mild acid hydrolysis, MALDI-TOF MS, ZIC-HILIC, ZIC-HILIC-ESI-MS
Comments, role: ECA(CYC), cyclic glycoforms were identified with different level of OAc groups. c[ECA(CYC)]4 was non O-acetylated in S. sonnei phase II, and E. coli R1, and for E. coli K12 +1-2OAc. c[ECA(CYC)]5-6 +3-4OAc, +2OAc and +1-5OAc respectively.
Related record ID(s): 10855
NCBI Taxonomy refs (TaxIDs): 624,
562,
316407
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There is only one chemically distinct structure:
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Gozdziewicz TK, Maciejewska A, Tsybulska A, Lugowski C, Lukasiewicz J
A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification
International Journal of Molecular Sciences 22(2) (2021)
701
|
-3)-a-D-Fucp4NAc-(1-4)-b-D-ManpNAcA-(1-4)-a-D-GlcpNAc6(%)Ac-(1- |
Show graphically |
Shigella sonnei PCM 1985
(Ancestor NCBI TaxID 624,
species name lookup)
Escherichia coli R1 F470
(Ancestor NCBI TaxID 562,
species name lookup)
Escherichia coli K12 W3110
(NCBI TaxID 316407,
species name lookup)
Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Shigella sonnei [ICD11:
XN9M9 ];
infection due to Escherichia coli [ICD11:
XN6P4 ]
The structure was elucidated in this paperNCBI PubMed ID: 33445776Publication DOI: 10.3390/ijms22020701Journal NLM ID: 101092791Publisher: 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 antigen expressed by enterobacteria. It is built by trisaccharide repeating units: →3)-α-D-Fucp4NAc-(1→4)-β-D-ManpNAcA-(1→4)-α-D-GlcpNAc-(1→ and occurs in three forms: as surface-bound linear polysaccharides linked to a phosphoglyceride (ECAPG) or lipopolysaccharide - endotoxin (ECALPS), and cyclic form (ECACYC). ECA maintains, outer membrane integrity, immunogenicity, and viability of enterobacteria. A supernatant obtained after LPS ultracentrifugation was reported as a source for ECA isolation, but it has never been assessed for detailed composition besides ECACYC. We used mild acid hydrolysis and gel filtration, or zwitterionic-hydrophilic interaction liquid (ZIC®HILIC) chromatography combined with mass spectrometry for purification, fractionation, and structural analysis of rough Shigella sonnei and Escherichia coli R1 and K12 crude LPS preparations. Presented work is the first report concerning complex characteristic of all ECA forms present in LPS-derived supernatants. We demonstrated high heterogeneity of the supernatant-derived ECA that contaminate LPS purified by ultracentrifugation. Not only previously reported O-acetylated tetrameric, pentameric, and hexameric ECACYC have been identified, but also devoid of lipid moiety linear ECA built from 7 to 11 repeating units. Described results were common for all selected strains. The origin of linear ECA is discussed against the current knowledge about ECAPG and ECALPS.
Lipopolysaccharide, LPS, enterobacterial common antigen, ECA, mass spectrometry, cyclic ECA, ECAPG
Structure type: polymer chemical repeating unit ; n=7,10,11
Location inside paper: abstract, Fig. 2, table 1
Trivial name: ECA(PG) linear form
Compound class: Enterobacterial common antigen
Contained glycoepitopes: IEDB_141807,IEDB_151531
Methods: gel filtration, ESI-MS, mild acid hydrolysis, MALDI-TOF MS, ZIC-HILIC, ZIC-HILIC-ESI-MS
Comments, role: linear form ECA of S. sonnei phase II contained 4–9 O-Ac groups per molecule, E. coli R1 - 0–5 O-Ac groups per molecule and E. coli K12 non O-acetylated.
Related record ID(s): 7992
NCBI Taxonomy refs (TaxIDs): 624,
562,
316407
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There is only one chemically distinct structure:
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