title: Determination of the interface of a large protein complex by transferred cross-saturation measurements.
abstract: In an earlier paper, it was shown that the cross-saturation method enables us to identify the contact residues of large protein complexes in a more rigorous manner than is possible using chemical shift perturbation and hydrogen-deuterium exchange experiments. However, there are limitations within the determination of the contact residues by the cross-saturation method, in that the method is difficult to apply to protein complexes with a molecular mass over 150 kDa and/or with weak binding, since the resonances originating from the complexes should be observed directly in the method. In the present work, to overcome these limitations, we carried out the cross-saturation measurements under conditions of a fast exchange between free and bound states on the NMR time-scale, and determined the contact residues of the complex of the B domain of protein A and intact IgG, which has a molecular mass of 164 kDa and shows weak binding.
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Tamiji
authors.initials: T
authors.lastname: Nakanishi
authors.forename: Mayumi
authors.initials: M
authors.lastname: Miyazawa
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Sakakura
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Terasawa
authors.forename: Hideo
authors.initials: H
authors.lastname: Takahashi
authors.forename: Ichio
authors.initials: I
authors.lastname: Shimada
mesh_terms: D000818:Animals; D001665:Binding Sites; D007074:Immunoglobulin G / Q000737:chemistry*; D066298:In Vitro Techniques; D008024:Ligands; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008958:Models, Molecular; D008970:Molecular Weight; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013205:Staphylococcal Protein A / Q000737:chemistry*; D020349:Surface Plasmon Resonance
chemical_list: D007074:Immunoglobulin G; D008024:Ligands; D046911:Macromolecular Substances; D013205:Staphylococcal Protein A