title: 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.
abstract: 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.
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II", Via Cinthia 4, 80126 Napoli, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Marchetti
authors.forename: Angeles
authors.initials: A
authors.lastname: Canales
authors.forename: Rosa
authors.initials: R
authors.lastname: Lanzetta
authors.forename: Inga
authors.initials: I
authors.lastname: Nilsson
authors.forename: Christian
authors.initials: C
authors.lastname: Vogel
authors.forename: Dana E
authors.initials: DE
authors.lastname: Reed
authors.forename: David P
authors.initials: DP
authors.lastname: Aucoin
authors.forename: Jesús
authors.initials: J
authors.lastname: Jiménez-Barbero
authors.forename: Antonio
authors.initials: A
authors.lastname: Molinaro
authors.forename: Alba
authors.initials: A
authors.lastname: Silipo
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D019117:Burkholderia / Q000737:chemistry*; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000737:chemistry; D008070:Lipopolysaccharides; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D019081:O Antigens / Q000737:chemistry*; D014312:Trisaccharides / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D004187:Disaccharides; D008070:Lipopolysaccharides; D019081:O Antigens; D014312:Trisaccharides
keywords: Burkholderia anthina; LPS-antibody interaction; NMR spectroscopy; molecular modeling