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Unraveling the Interaction between the LPS O‐Antigen of Burkholderia anthina and the 5D8 Monoclonal Antibody by Using a Multidisciplinary Chemical Approach, with Synthesis, NMR, and Molecular Modeling Methods
Author(s) -
Marchetti Roberta,
Canales Angeles,
Lanzetta Rosa,
Nilsson Inga,
Vogel Christian,
Reed Dana E.,
AuCoin David P.,
JiménezBarbero Jesús,
Molinaro Antonio,
Silipo Alba
Publication year - 2013
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201300225
Subject(s) - epitope , chemistry , two dimensional nuclear magnetic resonance spectroscopy , monoclonal antibody , nuclear magnetic resonance spectroscopy , trisaccharide , stereochemistry , carbohydrate conformation , molecular model , biochemistry , computational biology , antigen , antibody , biology , genetics
The interaction between the O‐chain from the lipopolysaccharide from Burkholderia anthina and a lipopolysaccharide‐specific monoclonal antibody (5D8) has been studied at high resolution by NMR spectroscopy. In particular, the 5D8‐bound epitope of the saccharide entity has been unraveled by a combination of saturation transfer difference (STD) and transferred NOESY (tr‐NOESY) experiments performed on the 5D8/polysaccharide complex. To dissect the fine details of the molecular recognition events, further experiments with simpler carbohydrate ligands were carried out. Thus, experiments were also performed with ad hoc synthesized trisaccharide and hexasaccharide O‐antigen repeating units. By using this multidisciplinary approach (chemical synthesis, NMR spectroscopy and molecular dynamics simulation), determination of the binding epitope and the contribution to the binding of the sugar units composing the O‐chain have been determined.