A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification
Enterobacterial common antigen (ECA) is a conserved antigen expressed by enterobacteria. It is built by trisaccharide repeating units: →3)-α-D-Fuc<i>p</i>4NAc-(1→4)-β-D-Man<i>p</i>NAcA-(1→4)-α-D-Glc<i>p</i>NAc-(1→ and occurs in three forms: as surface-bound linear...
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MDPI AG
2021-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/2/701 |
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| author | Tomasz K Gozdziewicz Anna Maciejewska Alona Tsybulska Czeslaw Lugowski Jolanta Lukasiewicz |
| author_facet | Tomasz K Gozdziewicz Anna Maciejewska Alona Tsybulska Czeslaw Lugowski Jolanta Lukasiewicz |
| author_sort | Tomasz K Gozdziewicz |
| collection | DOAJ |
| description | Enterobacterial common antigen (ECA) is a conserved antigen expressed by enterobacteria. It is built by trisaccharide repeating units: →3)-α-D-Fuc<i>p</i>4NAc-(1→4)-β-D-Man<i>p</i>NAcA-(1→4)-α-D-Glc<i>p</i>NAc-(1→ and occurs in three forms: as surface-bound linear polysaccharides linked to a phosphoglyceride (ECA<sub>PG</sub>) or lipopolysaccharide − endotoxin (ECA<sub>LPS</sub>), and cyclic form (ECA<sub>CYC</sub>). 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 ECA<sub>CYC</sub>. We used mild acid hydrolysis and gel filtration, or zwitterionic-hydrophilic interaction liquid (ZIC<sup>®</sup>HILIC) chromatography combined with mass spectrometry for purification, fractionation, and structural analysis of rough <i>Shigella sonnei</i> and <i>Escherichia coli</i> 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 <i>O</i>-acetylated tetrameric, pentameric, and hexameric ECA<sub>CYC</sub> 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 ECA<sub>PG</sub> and ECA<sub>LPS</sub>. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T05:02:15Z |
| publishDate | 2021-01-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9d22b7944e9341a3b6309238bb7fb84f2023-12-03T12:58:41ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-01-0122270110.3390/ijms22020701A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides PurificationTomasz K Gozdziewicz0Anna Maciejewska1Alona Tsybulska2Czeslaw Lugowski3Jolanta Lukasiewicz4Laboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, PolandLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, PolandLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, PolandLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, PolandLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, PolandEnterobacterial common antigen (ECA) is a conserved antigen expressed by enterobacteria. It is built by trisaccharide repeating units: →3)-α-D-Fuc<i>p</i>4NAc-(1→4)-β-D-Man<i>p</i>NAcA-(1→4)-α-D-Glc<i>p</i>NAc-(1→ and occurs in three forms: as surface-bound linear polysaccharides linked to a phosphoglyceride (ECA<sub>PG</sub>) or lipopolysaccharide − endotoxin (ECA<sub>LPS</sub>), and cyclic form (ECA<sub>CYC</sub>). 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 ECA<sub>CYC</sub>. We used mild acid hydrolysis and gel filtration, or zwitterionic-hydrophilic interaction liquid (ZIC<sup>®</sup>HILIC) chromatography combined with mass spectrometry for purification, fractionation, and structural analysis of rough <i>Shigella sonnei</i> and <i>Escherichia coli</i> 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 <i>O</i>-acetylated tetrameric, pentameric, and hexameric ECA<sub>CYC</sub> 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 ECA<sub>PG</sub> and ECA<sub>LPS</sub>.https://www.mdpi.com/1422-0067/22/2/701enterobacterial common antigenECAcyclic ECAECA<sub>PG</sub>lipopolysaccharideLPS |
| spellingShingle | Tomasz K Gozdziewicz Anna Maciejewska Alona Tsybulska Czeslaw Lugowski Jolanta Lukasiewicz A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification International Journal of Molecular Sciences enterobacterial common antigen ECA cyclic ECA ECA<sub>PG</sub> lipopolysaccharide LPS |
| title | A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification |
| title_full | A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification |
| title_fullStr | A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification |
| title_full_unstemmed | A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification |
| title_short | A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification |
| title_sort | new look at the enterobacterial common antigen forms obtained during rough lipopolysaccharides purification |
| topic | enterobacterial common antigen ECA cyclic ECA ECA<sub>PG</sub> lipopolysaccharide LPS |
| url | https://www.mdpi.com/1422-0067/22/2/701 |
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