Bivalent binding drives the formation of the Grb2-Gab1 signaling complex in a noncooperative manner

Caleb B. McDonald, Vikas Bhat, David C. Mikles, Brian J. Deegan, Kenneth L. Seldeen, Amjad Farooq

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2156-2173
Number of pages18
JournalFEBS Journal
Volume279
Issue number12
DOIs
StatePublished - Jun 2012

Keywords

  • intrinsic disorder
  • macromolecular assembly
  • multivalent binding
  • SH3-ligand interactions
  • zero cooperativity

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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