title: Surface plasmon resonance thermodynamic and kinetic analysis as a strategic tool in drug design. Distinct ways for phosphopeptides to plug into Src- and Grb2 SH2 domains.
abstract: Thermodynamic and kinetic studies of biomolecular interactions give insight into specificity of molecular recognition processes and advance rational drug design. Binding of phosphotyrosine (pY)-containing peptides to Src- and Grb2-SH2 domains was investigated using a surface plasmon resonance (SPR)-based method. This SPR assay yielded thermodynamic binding constants in solution, and the kinetic information contained in the SPR signal allowed kinetic analysis, which demonstrated distinct ways for pY ligands to interact with the SH2 domains. The results for binding to Src SH2 were consistent with sequestration of water molecules in the interface of the pYEEI peptide/Src SH2 complex. The results for a pYVNV peptide binding to Grb2 SH2 suggested a conformational change for Grb2 SH2 upon binding, which is not observed for Src SH2. Binding of a cyclic construct, allowing the pYVNV sequence in the bound conformation, did not have the expected entropy advantage. The results suggest an alternative binding mode for this construct, with the hydrophobic ring-closing part interacting with the protein. In all cases, except for full-length Grb2 protein, the affinity for the immobilized peptide at the SPR sensor and in solution was identical. This study demonstrates that SPR thermodynamic and kinetic analysis is a useful strategic tool in drug design.
authors.affiliation: Department of Medicinal Chemistry, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht, The Netherlands. n.j.demol@pharm.uu.nl
authors.forename: Nico J
authors.initials: NJ
authors.lastname: de Mol
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Dekker
authors.forename: Isabel
authors.initials: I
authors.lastname: Broutin
authors.forename: Marcel J E
authors.initials: MJ
authors.lastname: Fischer
authors.forename: Rob M J
authors.initials: RM
authors.lastname: Liskamp
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry*; D015195:Drug Design; D051380:GRB2 Adaptor Protein; D007700:Kinetics; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D015688:Oncogene Protein pp60(v-src) / Q000737:chemistry*; D010748:Phosphopeptides / Q000737:chemistry*; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D013816:Thermodynamics; D018909:src Homology Domains
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; D009842:Oligopeptides; D010748:Phosphopeptides; C487410:phosphotyrosyl-glutamyl-glutamyl-isoleucine; D015688:Oncogene Protein pp60(v-src)