Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Brucella abortus [ICD11:
XN7A8 
]
NCBI PubMed ID: 8576040Journal NLM ID: 2985120RPublisher: American Society for Microbiology
Correspondence: iMORiYON
MAiL2.CTi.UNAV.ES
Institutions: Departamento de Microbiologia, Clinica Universitaria y Facultad de Medicina, Universidad de Navarra, Pamplona, Spain, Programa de Investigacion en Enfermedades Tropicales (PIEt), Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica
Brucella native haptens (NHs) extracted with hot water from smooth (S)-type B. abortus and B. melitensis were purified to high levels of serological activity and compared with the polysaccharide obtained by acid hydrolysis (PS) of the S lipopolysaccharide (S-LPS). By 13C nuclear magnetic resonance analysis, NHs showed the spectrum of a homopolymer of a-1,2- or a-1,2- plus a-1,3-linked 4-formamido-4,6-dideoxy-D-mannose (N-formylperosamine) previously reported for the LPS O chain. However, while PS contained up to 0.6% 3-deoxy-D-manno-2-octulosonate, this LPS-core marker was absent from NH. High performance liquid chromatography and thin-layer chromatography showed heterogeneity in NH purified from whole cells but not in PS. By immunoprecipitation, polysaccharides indistinguishable from NH were demonstrated in extracts obtained with phenol-water, saline at 60 C, and ether-water treatments, and none of these treatments caused S-LPS hydrolysis detectable with antibodies to the O chain and lipid A. Two lines of evidence showed that NH was in the cell surface. First, NH became biotinylated when B. abortus live cells were labelled with biotin-hydrazide, and the examination of cell fractions and electron microscopy sections with streptavidin-peroxidase and streptavidin-coloidal gold, respectively, showed that labelling was extrinsic. Moreover, whereas only traces of NH were found in cytosols, the amount of NH was enriched in cell envelopes and in the outer membrane blebs spontaneously released by brucellae during growth. Interactions between NH and S-LPS were observed in crude cell extracts, and such interactions could be reconstituted by using purified NH and LPS. The results demonstrate that NH is not a hydrolytic product of S-LPS and suggest a model in which LPS-independent O-type polysaccharides (NH) are intertwined with the O chain in the outer membrane of S-type brucellae.
Lipopolysaccharide, LPS, characterization, polysaccharide, polysaccharides, Brucella, Brucella abortus, hapten, Brucella melitensis, membrane, outer membrane, native, Haptens, O-type, smooth
Structure type: homopolymer
Location inside paper: Table. 1, Brucella abortus 2308 fractions HW-5 and HW-6
Trivial name: perosamine homopolymer, perosamine
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_131172,IEDB_134281,IEDB_1397515,IEDB_2116320,IEDB_434547,IEDB_628715
Methods: gel filtration, NMR, SDS-PAGE, HPLC, immunoblotting, MAb studies, gel immunoprecipitation, HPTLC, extrinsic labelling
Biological activity: immunological data
Comments, role: NMR temperature: 296 and 309
Related record ID(s): 42, 221, 1236, 3181, 5252, 6859, 7225, 7463, 10088, 108433, 110473, 122777, 130189, 143514, 148072
NCBI Taxonomy refs (TaxIDs): 359391Reference(s) to other database(s): GTC:G55700WU, GlycomeDB:
3393, CCSD:
42651, CBank-STR:4424
Show glycosyltransferases
NMR conditions: in D2O at 303(C) K
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
4 Fo
aDRhap4N 101.65 78.05 69.35 52.96 68.72 17.99
1H NMR data:
missing...
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 |
|---|
| 4 | Fo | |
| | aDRhap4N | 101.65 | 78.05 | 69.35 | 52.96 | 68.72 | 17.99 |
|
There is only one chemically distinct structure:
Found 
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