title: Ligand binding by antibody IgE Lb4: assessment of binding site preferences using microcalorimetry, docking, and free energy simulations.
abstract: Antibody IgE Lb4 interacts favorably with a large number of different compounds. To improve the current understanding of the structural basis of this vast cross-reactivity, the binding of three dinitrophenyl (DNP) amino acids (DNP-alanine, DNP-glycine, and DNP-serine) is investigated in detail by means of docking and molecular dynamics free energy simulations. Experimental binding energies obtained by isothermal titration microcalorimetry are used to judge the results of the computational studies. For all three ligands, the docking procedure proposes two plausible subsites within the binding region formed by the antibody CDR loops. By subsequent molecular dynamics simulations and calculations of relative free energies of binding, one of these subsites, a tyrosine-surrounded pocket, is revealed as the preferred point of complexation. For this subsite, results consistent with experimental observations are obtained; DNP-glycine is found to bind better than DNP-serine, and this, in turn, is found to bind better than DNP-alanine. The suggested binding mode makes it possible to explain both the moderate binding affinity and the differences in binding energy among the three ligands.
authors.affiliation: Institute of General, Inorganic and Theoretical Chemistry, University of Innsbruck, A-6020 Innsbruck, Austria.
authors.forename: C A
authors.initials: CA
authors.lastname: Sotriffer
authors.forename: W
authors.initials: W
authors.lastname: Flader
authors.forename: A
authors.initials: A
authors.lastname: Cooper
authors.forename: B M
authors.initials: BM
authors.lastname: Rode
authors.forename: D S
authors.initials: DS
authors.lastname: Linthicum
authors.forename: K R
authors.initials: KR
authors.lastname: Liedl
authors.forename: J M
authors.initials: JM
authors.lastname: Varga
mesh_terms: D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002151:Calorimetry; D003429:Cross Reactions; D004136:Dinitrobenzenes / Q000276:immunology; D005998:Glycine / Q000031:analogs & derivatives / Q000276:immunology; D007073:Immunoglobulin E / Q000737:chemistry* / Q000378:metabolism*; D066298:In Vitro Techniques; D008024:Ligands; D008958:Models, Molecular; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000276:immunology; D011487:Protein Conformation; D012694:Serine / Q000031:analogs & derivatives / Q000276:immunology; D013816:Thermodynamics
chemical_list: D004136:Dinitrobenzenes; D008024:Ligands; C028545:N-(2,4-dinitrophenyl)alanine; D007073:Immunoglobulin E; D012694:Serine; D010649:Phenylalanine; D005998:Glycine