Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
NCBI PubMed ID: 12230919Journal NLM ID: 9433350Publisher: Maney Publishing
Correspondence: hbrade
fz-borstel.de
Institutions: Divisions of 1Biochemical and Medical Microbiology and Biophysics, Center for Medicine and Biosciences, Research Center Borstel, Borstel, Germany, Institute of Chemistry, University of Agricultural Sciences, Vienna, Austria
We report a novel strategy for the preparation of neoglycoconjugates of oligosaccharides which are obtained after complete deacylation of bacterial deep rough lipopolysaccharides (LPS) isolated from recombinant Escherichia coli bacteria synthesizing a Kdo di-[a-Kdo-(24)-a-Kdo-(2] and a Kdo trisaccharide [a-Kdo-(28)-a-Kdo-(24)-a-Kdo-(2] of Re-type and chlamydial LPS, respectively. Unlike acylated LPS, such oligosaccharides can be obtained in pure form and thus lead to well-defined neoglycoconjugates. Cleavage of the 1-phosphate of the lipid A moiety by alkaline phosphatase treatment leads to a free reducing glucosamine which can be further reacted with allylamine. After reductive amination, spacer elongation of the allyl group with cysteamine and activation with thiophosgene, the ligands were reacted with BSA. We have compared the immunological reactivity of such defined neoglycoconjugates obtained from natural sources with those obtained by chemical synthesis and report that such neoglycoconjugates are immunogenic and well suited as antigens for the study of epitope specificities of monoclonal antibodies. In addition, we have compared these conjugates with those in which ligands were coupled by glutardialdehyde to BSA. Our approach proved to be superior since the latter led upon immunization of mice to a relatively high percentage of antibodies that reacted with glutardialdehyde derivatized BSA without the carbohydrate ligand. This was not the case for cysteamine-spacered ligands coupled via their isothiocyanate-derivatives.
Lipopolysaccharide, synthesis, lipopolysaccharides, LPS, oligosaccharide, Bacterial, trisaccharide, group, form, Escherichia, Escherichia coli, Kdo, lipid, lipid A, Oligosaccharides, bacteria, mutant, lead, alkaline, rough, treatment, deacylated, deacylation, reducing, preparation, recombinant, acylated, Glucosamine, spacer, phosphatase, free, cleavage, chlamydial, alkaline phosphatase, amination, reductive, neoglycoconjugate, allyl, deacylated LPS, elongation, neoglycocnjugate
Structure type: oligomer
Location inside paper: p. 297, Table 1
Contained glycoepitopes: IEDB_130650,IEDB_130657,IEDB_130658,IEDB_130659,IEDB_141807,IEDB_151531
Methods: NMR, MS
Biological activity: serological studies with monoclonal antibodies
Enzymes that release or process the structure: alcaline phosphatase
Synthetic data: chemoenzymatic
Comments, role: neoglycoconjugates from deacylated deep rough lipopolysaccharides of Escherichia coli
Related record ID(s): 3724, 3806, 3807, 3808, 3809, 3810, 3811
NCBI Taxonomy refs (TaxIDs): 562Reference(s) to other database(s): GTC:G70367QE
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
report error