Taxonomic group: bacteria /
NCBI PubMed ID: 10350463Journal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: thomas.peters
cheme.muluebeck.de
Institutions: Institut fur Chemie, Medizinische Universitat Lubeck, Ratzeburger Allee 160, D-23538 Lubeck, Germany, Instituto de Quimica Organica General, CSIC, Juan de la Cierva 3, E-28006 Madrid, Spain, Institut fur Chemie der Universitat fur Bodenkultur Wien, A-1190 Wien, Austria, Forschungszentrum Borstel, Zentrum fur Medizin und Biowissenschaften, Borstel, Germany
The recognition reactions between a synthetic disaccharide α-Kdo-(2→4)-α-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 α-Kdo-(2→4)-α-Kdo-(2→O)-allyl and α-Kdo-(2→8)-α-Kdo-(2→O)-allyl with similar affinities, whereas mAb S25-2 binds to the disaccharide α-Kdo-(2→8)-α-Kdo-(2→O)-allyl with an approximately 10-fold higher affinity than to the disaccharide α-Kdo-(2→4)-α-Kdo-(2→O)-allyl. Compared to S25-2, S23-24 binds to α-Kdo-(2→4)-α-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 α-(2→4)-linked disaccharide that is not highly populated in solution. S23-24 recognizes two conformations of α-Kdo-(2→4)-α-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.
Lipopolysaccharide, NMR, conformation, Bacterial, structural, epitope, epitopes, disaccharide, Synthetic
Structure type: oligomer ; 597.6
Location inside paper: disaccharide 1, 6452
Contained glycoepitopes: IEDB_130650,IEDB_130659
Methods: NMR, trNOESY, trROESY, QUIET-trNOESY, MINSY
Biological activity: generation of monoclonal antibodies (IgG1 type); binding of mAbs to disaccharide 1 and 2
Comments, role: synthetic disaccharide (dipotassium salt, monohydrate)
3D data: conformation data
Related record ID(s): 565, 3186, 4142, 116696
NCBI Taxonomy refs (TaxIDs): 2Reference(s) to other database(s): GlycomeDB:
5576
Show glycosyltransferases
NMR conditions: in D2O at 310(H) K
[as TSV]
13C NMR data:
missing...
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6 H7 H8
1,4 aXKdop - - 1.87-2.24 4.18 4.14 3.71 4.08 3.84-4.07
1 aXKdop - - 2.02-2.10 4.24 4.18 3.65 4.04 3.71-4.02
Allyl
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 | H7 | H8 |
|---|
| 1,4 | aXKdop |
|
| 1.87
2.24 | 4.18 | 4.14 | 3.71 | 4.08 | 3.84
4.07 |
| 1 | aXKdop |
|
| 2.02
2.10 | 4.24 | 4.18 | 3.65 | 4.04 | 3.71
4.02 |
| | Allyl | |
|
There is only one chemically distinct structure:
Taxonomic group: bacteria /
NCBI PubMed ID: 10350463Journal NLM ID: 0370623Publisher: American Chemical Society
Correspondence: thomas.peters
cheme.muluebeck.de
Institutions: Institut fur Chemie, Medizinische Universitat Lubeck, Ratzeburger Allee 160, D-23538 Lubeck, Germany, Instituto de Quimica Organica General, CSIC, Juan de la Cierva 3, E-28006 Madrid, Spain, Institut fur Chemie der Universitat fur Bodenkultur Wien, A-1190 Wien, Austria, Forschungszentrum Borstel, Zentrum fur Medizin und Biowissenschaften, Borstel, Germany
The recognition reactions between a synthetic disaccharide α-Kdo-(2→4)-α-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 α-Kdo-(2→4)-α-Kdo-(2→O)-allyl and α-Kdo-(2→8)-α-Kdo-(2→O)-allyl with similar affinities, whereas mAb S25-2 binds to the disaccharide α-Kdo-(2→8)-α-Kdo-(2→O)-allyl with an approximately 10-fold higher affinity than to the disaccharide α-Kdo-(2→4)-α-Kdo-(2→O)-allyl. Compared to S25-2, S23-24 binds to α-Kdo-(2→4)-α-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 α-(2→4)-linked disaccharide that is not highly populated in solution. S23-24 recognizes two conformations of α-Kdo-(2→4)-α-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.
Lipopolysaccharide, NMR, conformation, Bacterial, structural, epitope, epitopes, disaccharide, Synthetic
Structure type: oligomer ; 597.6
Location inside paper: disaccharide 2, 6452
Contained glycoepitopes: IEDB_130650,IEDB_130658
Methods: NMR, trNOESY, trROESY, QUIET-trNOESY, MINSY
Biological activity: generation of monoclonal antibodies (IgG1 type); binding of mAbs to disaccharide 1 and 2
Comments, role: synthetic disaccharide (dipotassium salt, monohydrate)
3D data: conformation data
Related record ID(s): 496, 3235, 3880, 4143, 4921, 22428, 116695
NCBI Taxonomy refs (TaxIDs): 2Reference(s) to other database(s): GlycomeDB:
5769
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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