NMR Experiments Reveal Distinct Antibody-Bound Conformations of a Synthetic Disaccharide Representing a General Structural Element of Bacterial Lipopolysaccharide Epitopes
| ISSN: | 1520-4995 0006-2960 |
|---|---|
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c27c4e38543345e9b3024c0209bd3e08 https://doi.org/10.1021/bi982984z |
| Přírůstkové číslo: | edsair.doi.dedup.....c27c4e38543345e9b3024c0209bd3e08 |
| Autor: | Jesús Jiménez-Barbero, Lore Brade, Thomas Haselhorst, Tobias Sokolowski, Juan-Félix Espinosa, Thomas Peters, Helmut Brade, Paul Kosma |
| Rok vydání: | 1999 |
| Předmět: |
Lipopolysaccharides
Aqueous solution Stereochemistry Ligand Disaccharide Antibodies Monoclonal Carbohydrate Disaccharides Ligands Antibodies Bacterial Biochemistry Affinities Epitope Epitopes chemistry.chemical_compound chemistry Carbohydrate Conformation Humans Binding Sites Antibody Carbohydrate conformation Chlamydia Binding site Nuclear Magnetic Resonance Biomolecular |
| Zdroj: | Biochemistry. 38:6449-6459 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|