Changes in structural dynamics of the Grb2 adaptor protein upon binding of phosphotyrosine ligand to its SH2 domain.
Growth factor receptor-bound protein 2 (Grb2) is an extensively studied adaptor protein involved in cell signaling. Grb2 is a highly flexible protein composed of a single SH2 domain flanked by two SH3 domains. Here we report on the structural dynamic effects upon interaction of a phosphopeptide ligand derived from the recognition sequence of the Shc adaptor protein with (i) the isolated SH2 domain of Grb2 (Grb2 SH2) and (ii) the full-length Grb2 protein. From kinetic studies using surface plasmon resonance, it was deduced that a conformation change occurred in the SH2 protein as well as the full-length Grb2 after binding. Measurements of hydrogen/deuterium exchange (HDX) in the isolated SH2 domain and full-length Grb2 protein as monitored by electrospray mass spectrometry, showed that binding reduces the overall flexibility of the proteins, possibly via slightly different mechanisms for the single SH2 domain and the full-length Grb2 protein.
de Mol
Nico J
NJ
Department of Medicinal Chemistry, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, Sorbonnelaan 16, 3508 TB Utrecht, The Netherlands. N.J.deMol@pharm.uu.nl
Catalina
M Isabel
MI
Fischer
Marcel J E
MJ
Broutin
Isabelle
I
Maier
Claudia S
CS
Heck
Albert J R
AJ
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D041961:Deuterium Exchange Measurement; D051380:GRB2 Adaptor Protein; D007700:Kinetics; D008024:Ligands; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D019000:Phosphotyrosine / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D055765:Shc Signaling Adaptor Proteins; D021241:Spectrometry, Mass, Electrospray Ionization; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D020349:Surface Plasmon Resonance; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D051380:GRB2 Adaptor Protein; C495949:Grb2 protein, mouse; D008024:Ligands; D010446:Peptide Fragments; D011506:Proteins; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D019000:Phosphotyrosine