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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 paper
NCBI PubMed ID: 35653593
Publication DOI: 10.1021/acsinfecdis.2c00183
Journal NLM ID: 101654580
Publisher: 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: oligomer
Location inside paper: p. 1337, Fig. 1, table 1, OS1
Trivial name: A-band polysaccharide (A-PS)
Compound class: CPA
Contained glycoepitopes: IEDB_114707,IEDB_130701,IEDB_1394181,IEDB_144983,IEDB_152206,IEDB_983930,SB_44,SB_67,SB_72

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: oligosaccharide OS1 (the reducing end is 1-deuterated erythritol) was obtained by periodate oxidation of the A-PS with subsequent NaBD4 reduction and mild hydrolysis.

Related record ID(s): 8437, 20856, 20858, 20859, 20860
NCBI Taxonomy refs (TaxIDs): 208964
Show glycosyltransferases

NMR conditions: in D2O at 298 K [as TSV]

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
2,4,3	Me	57.5
2,4	aDRhap	102.4	67.4	81.7	79.1	69.0	18.3
2,4,4,3	Me	57.5
2,4,4	aDRhap	102.4	67.3	80.6	71.8	70.6	17.7
2,3	Me	57.5
2	aDManp	99.8	67.3	81.9	73.8	72.9	67.1
	x?Ery-ol	59.7	78.5	72.3	63.7


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
2,4,3	Me	3.46
2,4	aDRhap	5.15	4.21	3.58	3.59	3.89	1.33
2,4,4,3	Me	3.46
2,4,4	aDRhap	5.16	4.23	3.43	3.50	3.80	1.29
2,3	Me	3.46
2	aDManp	5.12	4.24	3.67	3.83	3.83	3.78-3.85
	x?Ery-ol	3.75-3.87	3.79	3.90	3.63-3.78


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
2,4,3	Me	57.5/3.46
2,4	aDRhap	102.4/5.15	67.4/4.21	81.7/3.58	79.1/3.59	69.0/3.89	18.3/1.33
2,4,4,3	Me	57.5/3.46
2,4,4	aDRhap	102.4/5.16	67.3/4.23	80.6/3.43	71.8/3.50	70.6/3.80	17.7/1.29
2,3	Me	57.5/3.46
2	aDManp	99.8/5.12	67.3/4.24	81.9/3.67	73.8/3.83	72.9/3.83	67.1/3.78-3.85
	x?Ery-ol	59.7/3.75-3.87	78.5/3.79	72.3/3.90	63.7/3.63-3.78

¹H NMR data:

Linkage	Residue	H1	H2	H3	H4	H5	H6
2,4,3	Me	3.46
2,4	aDRhap	5.15	4.21	3.58	3.59	3.89	1.33
2,4,4,3	Me	3.46
2,4,4	aDRhap	5.16	4.23	3.43	3.50	3.80	1.29
2,3	Me	3.46
2	aDManp	5.12	4.24	3.67	3.83	3.83	3.78 3.85
	x?Ery-ol	3.75 3.87	3.79	3.90	3.63 3.78

¹³C NMR data:

Linkage	Residue	C1	C2	C3	C4	C5	C6
2,4,3	Me	57.5
2,4	aDRhap	102.4	67.4	81.7	79.1	69.0	18.3
2,4,4,3	Me	57.5
2,4,4	aDRhap	102.4	67.3	80.6	71.8	70.6	17.7
2,3	Me	57.5
2	aDManp	99.8	67.3	81.9	73.8	72.9	67.1
	x?Ery-ol	59.7	78.5	72.3	63.7

There are 2 chemically distinct structures. Please, select:

CO[C@@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H](C)O[C@H](O[C@@H]3[C@@H](C)O[C@H](O[C@@H]4[C@@H](C)O[C@H](O[C@@H]5[C@@H](C)O[C@H](O[C@@H]6[C@@H](CO)O[C@H](O[C@@H](CO)[C@H](O)CO)[C@@H](O)[C@H]6OC)[C@@H](O)[C@H]5OC)[C@@H](O)[C@H]4OC)[C@@H](O)[C@H]3OC)[C@@H](O)[C@H]2OC)O[C@H](C)[C@H]1O

SVG MW SMILES 3D mol Ionize