Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: cystic fibrosis (CF) [ICD11:
CA25 
];
infection due to Pseudomonas aeruginosa [ICD11:
XN5L6 ]
The structure was elucidated in this paperNCBI PubMed ID: 35653593Publication DOI: 10.1021/acsinfecdis.2c00183Journal NLM ID: 101654580Publisher: Washington, DC: American Chemical Society
Correspondence: A.D. Cox <Andrew.Cox
nrc-cnrc.gc.ca>
Institutions: Vaccine and Emerging Infections Research, Human Health Therapeutics Research Centre, National Research Council, Ottawa, Ontario K1A 0R6, Canada
Pseudomonas aeruginosa produces a variety of cell surface glycans. Previous studies identified a common polysaccharide (PS) antigen often termed A-band PS that was composed of a neutral d-rhamnan trisaccharide repeating unit as a relatively conserved cell surface carbohydrate. However, nuclear magnetic resonance (NMR) spectra and chemical analysis of A-PS preparations showed the presence of several additional components. Here, we report the characterization of the carbohydrate component responsible for these signals. The carbohydrate antigen consists of an immunogenic methylated rhamnan oligosaccharide at the nonreducing end of the A-band PS. Initial studies performed with the isolated antigen permitted the production of conjugates that were used to immunize mice and rabbits and generate monoclonal and polyclonal antibodies. The polyclonal antibodies were able to recognize the majority of P. aeruginosa strains in our collection, and three monoclonal antibodies were generated, one of which was able to recognize and facilitate opsonophagocytic killing of a majority of P. aeruginosa strains. This monoclonal antibody was able to recognize all P. aeruginosa strains in our collection that includes clinical and serotype strains. Synthetic oligosaccharides (mono- to pentasaccharides) representing the terminal 3-O-methyl d-rhamnan were prepared, and the trisaccharide was identified as the antigenic determinant required to effectively mimic the natural antigen recognized by the broadly cross-reactive monoclonal antibody. These data suggest that there is considerable promise in this antigen as a vaccine or therapeutic target.
Lipopolysaccharide, Pseudomonas aeruginosa, monoclonal antibody, glycoconjugate vaccine, common polysaccharide antigen, methylrhamnan
Structure type: polymer chemical repeating unit
Location inside paper: p. 1337, Fig. 1, A-PS repeating unit
Trivial name: common antigen, common polysaccharide antigen, A band polysaccharide, D-rhamnan, repeat unit of the lipopolysaccharide side-chain, A-band polysaccharide (A-PS), A-band polysaccharide
Compound class: O-polysaccharide, O-antigen, LPS, enterobacterial common antigen, CPA
Contained glycoepitopes: IEDB_1394181,IEDB_144831,IEDB_145005,IEDB_145006,IEDB_145010
Methods: 13C NMR, 1H NMR, periodate oxidation, NMR-2D, GC-MS, ELISA, ESI-MS, anion-exchange chromatography, MALDI-MS, chemical synthesis, HPLC, serum bactericidal assays, gel chromatography, SEC, SPR, immunization, conjugation, alkaline treatment, opsonophagocytic assay, isolation, mAb epitope mapping
Comments, role: published polymerization frame was shifted for conformity with other records
Related record ID(s): 20856, 20857, 20858, 20859, 20860
NCBI Taxonomy refs (TaxIDs): 208964Reference(s) to other database(s): GTC:G80135ZC, GlycomeDB:
25893
Show glycosyltransferases
NMR conditions: in D2O at 313 K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
2,3 aDRhap 103.5 71.2 79.1 72.8 70.5 18.0
2 aDRhap 103.3 71.2 79.3 72.8 70.7 18.0
aDRhap 102.1 79.4 71.2 73.6 70.5 18.0
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
2,3 aDRhap 5.05 4.15 3.91 3.58 3.89 1.32
2 aDRhap 4.97 4.17 3.85 3.58 3.77 1.29
aDRhap 5.21 4.09 3.97 3.52 3.85 1.32
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
2,3 aDRhap 103.5/5.05 71.2/4.15 79.1/3.91 72.8/3.58 70.5/3.89 18.0/1.32
2 aDRhap 103.3/4.97 71.2/4.17 79.3/3.85 72.8/3.58 70.7/3.77 18.0/1.29
aDRhap 102.1/5.21 79.4/4.09 71.2/3.97 73.6/3.52 70.5/3.85 18.0/1.32
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 |
|---|
| 2,3 | aDRhap | 5.05 | 4.15 | 3.91 | 3.58 | 3.89 | 1.32 |
| 2 | aDRhap | 4.97 | 4.17 | 3.85 | 3.58 | 3.77 | 1.29 |
| | aDRhap | 5.21 | 4.09 | 3.97 | 3.52 | 3.85 | 1.32 |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 |
|---|
| 2,3 | aDRhap | 103.5 | 71.2 | 79.1 | 72.8 | 70.5 | 18.0 |
| 2 | aDRhap | 103.3 | 71.2 | 79.3 | 72.8 | 70.7 | 18.0 |
| | aDRhap | 102.1 | 79.4 | 71.2 | 73.6 | 70.5 | 18.0 |
|
There is only one chemically distinct structure:
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