Abstract
Growth factor receptor-bound protein 2 (Grb2) is a small adapter protein composed of a single SH2 domain flanked by two SH3 domains. The N-terminal SH3 (nSH3) domain of Grb2 binds a proline-rich region present in the guanine nucleotide releasing factor, son of sevenless (Sos). Using NMR relaxation dispersion and chemical shift analysis methods, we investigated the conformational change of the Sos-derived proline-rich peptide during the transition between the free and Grb2 nSH3-bound states. The chemical shift analysis revealed that the peptide does not present a fully random conformation but has a relatively rigid structure. The relaxation dispersion analysis detected conformational exchange of several residues of the peptide upon binding to Grb2 nSH3.
Highlights
Growth factor receptor-bound protein 2 (Grb2) is a small adapter protein composed of a single SH2 domain flanked by two Src homology 3 (SH3) domains
Using NMR relaxation dispersion and chemical shift analysis methods, we investigated the conformational change of the son of sevenless (Sos)-derived proline-rich peptide during the transition between the free and Grb[2] N-terminal SH3 (nSH3)-bound states
Src homology 3 (SH3) domains are small protein modules consisting of approximately 60 amino acids that occur in cytoplasmic proteins, tyrosine kinases, and cytoskeletal proteins known to play important roles in signal transduction pathways[1]
Summary
Growth factor receptor-bound protein 2 (Grb2) is a small adapter protein composed of a single SH2 domain flanked by two SH3 domains. The N-terminal SH3 (nSH3) domain of Grb[2] binds a proline-rich region present in the guanine nucleotide releasing factor, son of sevenless (Sos). Using NMR relaxation dispersion and chemical shift analysis methods, we investigated the conformational change of the Sos-derived proline-rich peptide during the transition between the free and Grb[2] nSH3-bound states. Many structures of SH3 domains complexed with their specific ligand peptides have been solved by X-ray crystallography and NMR spectroscopy These structural studies have revealed that proline-rich peptides adopt a polyproline type II (PPII) helix on the SH3 domain[3,4,5,6,7]. We present here a 13Ca NMR relaxation dispersion analysis of the VPP peptide in the transition between free and bound states. Isotope-enriched samples are commonly used for relaxation dispersion measurements, Peng et al have demonstrated 13C relaxation dispersion at natural abundance levels for ligand peptides during protein binding[15,16], which has provided a comprehensive analysis of ms-ms conformational dynamics related to binding of the ligands
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