title: Solution structure of the Grb2 SH2 domain complexed with a high-affinity inhibitor.
abstract: The solution structure of the growth factor receptor-bound protein 2 (Grb2) SH2 domain complexed with a high-affinity inhibitor containing a non-phosphorus phosphate mimetic within a macrocyclic platform was determined by nuclear magnetic resonance (NMR) spectroscopy. Unambiguous assignments of the bound inhibitor and intermolecular NOEs between the Grb2 SH2 domain and the inhibitor was accomplished using perdeuterated Grb2 SH2 protein. The well-defined solution structure of the complex was obtained and compared to those by X-ray crystallography. Since the crystal structure of the Grb2 SH2 domain formed a domain-swapped dimer and several inhibitors were bound to a hinge region, there were appreciable differences between the solution and crystal structures. Based on the binding interactions between the inhibitor and the Grb2 SH2 domain in solution, we proposed a design of second-generation inhibitors that could be expected to have higher affinity.
authors.affiliation: Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, N12 W6, Sapporo, 060-0812, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Ogura
authors.forename: Takanori
authors.initials: T
authors.lastname: Shiga
authors.forename: Masashi
authors.initials: M
authors.lastname: Yokochi
authors.forename: Satoru
authors.initials: S
authors.lastname: Yuzawa
authors.forename: Terrence R
authors.initials: TR
authors.lastname: Burke
authors.forename: Fuyuhiko
authors.initials: F
authors.lastname: Inagaki
mesh_terms: D001665:Binding Sites; D032701:Biomimetics; D002850:Chromatography, Gel; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D051380:GRB2 Adaptor Protein / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010456:Peptides, Cyclic / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D013329:Structure-Activity Relationship; D018909:src Homology Domains
chemical_list: D051380:GRB2 Adaptor Protein; D010456:Peptides, Cyclic; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase