title: Bivalent binding drives the formation of the Grb2-Gab1 signaling complex in a noncooperative manner.
abstract: Although the growth factor receptor binder 2 (Grb2)-Grb2-associated binder (Gab)1 macromolecular complex mediates a multitude of cellular signaling cascades, the molecular basis of its assembly has hitherto remained largely elusive. Herein, using an array of biophysical techniques, we show that, whereas Grb2 exists in a monomer-dimer equilibrium, the proline-rich (PR) domain of Gab1 is a monomer in solution. Of particular interest is the observation that although the PR domain appears to be structurally disordered, it nonetheless adopts a more or less compact conformation reminiscent of natively folded globular proteins. Importantly, the structurally flexible conformation of the PR domain appears to facilitate the binding of Gab1 to Grb2 with a 1:2 stoichiometry. More specifically, the formation of the Grb2-Gab1 signaling complex is driven via a bivalent interaction through the binding of the C-terminal homology 3 (cSH3) domain within each monomer of Grb2 homodimer to two distinct RXXK motifs, herein designated G1 and G2, located within the PR domain of Gab1. Strikingly, in spite of the key role of bivalency in driving this macromolecular assembly, the cSH3 domains bind to the G1 and G2 motifs in an independent manner with zero cooperativity. Taken together, our findings shed new light on the physicochemical forces driving the assembly of a key macromolecular signaling complex that is relevant to cellular health and disease.
authors.affiliation: Department of Biochemistry & Molecular Biology and the USylvester Braman Family Breast Cancer Institute, Leonard Miller School of Medicine, University of Miami, FL 33136, USA.
authors.forename: Caleb B
authors.initials: CB
authors.lastname: McDonald
authors.forename: Vikas
authors.initials: V
authors.lastname: Bhat
authors.forename: David C
authors.initials: DC
authors.lastname: Mikles
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Deegan
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: Seldeen
authors.forename: Amjad
authors.initials: A
authors.lastname: Farooq
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051380:GRB2 Adaptor Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D011485:Protein Binding; D017433:Protein Structure, Secondary
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C098587:GAB1 protein, human; D051380:GRB2 Adaptor Protein