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Selected Publication:

Haselhorst, T; Espinosa, JF; Jiménez-Barbero, J; Sokolowski, T; Kosma, P; Brade, H; Brade, L; Peters, T.
(1999): NMR experiments reveal distinct antibody-bound conformations of a synthetic disaccharide representing a general structural element of bacterial lipopolysaccharide epitopes.
Biochemistry. 1999; 38(20):6449-6459 FullText FullText_BOKU

Abstract:
The recognition reactions between a synthetic disaccharide alpha-Kdo-(2-->4)-alpha-Kdo-(2-->O)-allyl and two monoclonal antibodies (mAbs) were studied by NMR, yielding two distinct bound conformations of the carbohydrate ligand. One mAb, S23-24, recognizes the disaccharides alpha-Kdo(2-->4)-alpha-Kdo-(2-->O)-allyl and alpha-Kdo-(2-->8)-alpha-Kdo-(2-->O)-allyl with similar affinities, whereas mAb S25-2 binds to the disaccharide alpha-Kdo-(2-->8)-alpha-Kdo-(2-->O)-allyl with an approximately 10-fold higher affinity than to the disaccharide alpha-Kdo-(2-->4)-alpha-Kdo-(2-->O)-allyl. Compared to S25-2, S23-24 binds to alpha-Kdo-(2-->4)-alpha Kdo-(2-->O)-allyl with an approximately 50-fold increased affinity. We used NMR experiments that are based on the transferred NOE effect, specifically, trNOESY, trROESY, QUIET-trNOESY, and MINSY experiments, to show that the (2-->8)-specific mAb, S25-2, stabilizes a conformation of the alpha-(2-->4)-linked disaccharide that is not highly populated in solution. S23-24 recognizes two conformations of alpha-Kdo-(2-->4)-alpha-Kdo-(2-->O)-allyl, one that is highly populated in aqueous solution and another conformation that is similar to the one bound by S25-2. This is the first example where it is experimentally shown that a carbohydrate ligand may adopt different bioactive conformations upon interaction with mAbs with different fine specificities. Our NMR studies indicate that a careful examination of spin diffusion is critical for the analysis of bioactive conformations of carbohydrate ligands.
Authors BOKU Wien:
Kosma Paul
Find related publications in this database (using NML MeSH Indexing)
Antibodies, Bacterial - chemistry
Antibodies, Monoclonal - chemistry
Binding Sites, Antibody - chemistry
Carbohydrate Conformation - chemistry
Chlamydia - immunology
Disaccharides - chemical synthesis
Epitopes - chemistry
Humans - chemistry
Ligands - chemistry
Lipopolysaccharides - chemistry
Nuclear Magnetic Resonance, Biomolecular - methods



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