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.

Structural Basis of Binding by Cyclic Nonphosphorylated Peptide Antagonists of Grb7 Implicated in Breast Cancer Progression

“…Besides the YXN motif interactions, the positioning of an aromatic residue against the βD6 side chain (as seen for F2 in the current structure) has similarly been observed for ligands bound to the Grb2-SH2 domain. 38,39 The G7-18NATE interaction with the Grb7-SH2 domain is, however, unlike that reported for the one other ligand-bound structure of the Grb7-SH2 domain. The structure of a 10-residue phosphorylated peptide, representing the pY1339 target site of the erbB-2 receptor, bound to the Grb7-SH2 domain has been solved using NMR spectroscopy 37 (PDB IDs: 1MW4 and 2L4K).…”

An NMR strategy for fragment-based ligand screening utilizing a paramagnetic lanthanide probe

“…The backbone amide signals of LBT-Grb2 containing Lu 3+ were assigned based on the assignments reported previously (Wang et al 1996; Thornton et al 1996; Ogura et al 2008). 1 H– 15 N HSQC spectra of the 15 N-labeled LBT-Grb2 were recorded in the presence of 1 equivalent of lanthanide ions: Lu 3+ , Tb 3+ , Dy 3+ , Er 3+ and Tm 3+ (Fig.…”

“…Conformer 1 of the family of NMR structures of Grb2 SH2 (1x0n.pdb, Ogura et al 2008) was used for the tensor fit.…”

The Molecular Basis of Phospholipase D2-Induced Chemotaxis: Elucidation of Differential Pathways in Macrophages and Fibroblasts

“…PLD was found to be elevated in cells that have been transformed by several oncogenes, including v-Src, v-Ras, v-Raf, and v-Fps, leading to the belief that there is a chronic turnover of PLD-dependent phosphatidylcholine (44). Increased amounts of Grb2 have been correlated with formation of tumors in the liver of mice as well as in human breast cancer cells (46), possibly leading to the ability to metastasize. Both cell lines studied were able to undergo chemotaxis through a permeable membrane in response to CSF-1 or EGF.…”