A novel NMR method for determining the interfaces of large protein-protein complexes.
Identification of the interfaces of large (Mr > 50,000) protein-protein complexes in solution by high resolution NMR has typically been achieved using experiments involving chemical shift perturbation and/or hydrogen-deuterium exchange of the main chain amide groups of the proteins. Interfaces identified using these techniques, however, are not always identical to those revealed using X-ray crystallography. In order to identify the contact residues in a large protein-protein complex more accurately, we developed a novel NMR method that uses cross-saturation phenomena in combination with TROSY detection in an optimally deuterium labeled system.
Takahashi
H
H
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Nakanishi
T
T
Kami
K
K
Arata
Y
Y
Shimada
I
I
D001665:Binding Sites; D018360:Crystallography, X-Ray; D003903:Deuterium / Q000378:metabolism; D004058:Diffusion; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry* / Q000378:metabolism*; D007553:Isotope Labeling; D046911:Macromolecular Substances; D008958:Models, Molecular; D008970:Molecular Weight; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011522:Protons; D015203:Reproducibility of Results; D012996:Solutions; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000378:metabolism*; D013211:Staphylococcus aureus
D007141:Immunoglobulin Fc Fragments; D046911:Macromolecular Substances; D011522:Protons; D012996:Solutions; D013205:Staphylococcal Protein A; D003903:Deuterium