title: Conformational recognition of an intrinsically disordered protein.
abstract: There is a growing interest in understanding the properties of intrinsically disordered proteins (IDPs); however, the characterization of these states remains an open challenge. IDPs appear to have functional roles that diverge from those of folded proteins and revolve around their ability to act as hubs for protein-protein interactions. To gain a better understanding of the modes of binding of IDPs, we combined statistical mechanics, calorimetry, and NMR spectroscopy to investigate the recognition and binding of a fragment from the disordered protein Gab2 by the growth factor receptor-bound protein 2 (Grb2), a key interaction for normal cell signaling and cancer development. Structural ensemble refinement by NMR chemical shifts, thermodynamics measurements, and analysis of point mutations indicated that the population of preexisting bound conformations in the free-state ensemble of Gab2 is an essential determinant for recognition and binding by Grb2. A key role was found for transient polyproline II (PPII) structures and extended conformations. Our findings are likely to have very general implications for the biological behavior of IDPs in light of the evidence that a large fraction of these proteins possess a specific propensity to form PPII and to adopt conformations that are more extended than the typical random-coil states.
authors.affiliation: Department of Life Sciences, Imperial College London, London, UK.
authors.forename: James M
authors.initials: JM
authors.lastname: Krieger
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge, UK.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Fusco
authors.affiliation: Department of Oncology, University of Oxford, Oxford, UK; Institute of Molecular Medicine, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Lewitzky
authors.affiliation: Department of Oncology, University of Oxford, Oxford, UK.
authors.forename: Philip C
authors.initials: PC
authors.lastname: Simister
authors.affiliation: Department of Life Sciences, Imperial College London, London, UK.
authors.forename: Jan
authors.initials: J
authors.lastname: Marchant
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge, UK.
authors.forename: Carlo
authors.initials: C
authors.lastname: Camilloni
authors.affiliation: Department of Oncology, University of Oxford, Oxford, UK; Institute of Molecular Medicine, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Stephan M
authors.initials: SM
authors.lastname: Feller
authors.affiliation: Department of Life Sciences, Imperial College London, London, UK. Electronic address: adesimon@imperial.ac.uk.
authors.forename: Alfonso
authors.initials: A
authors.lastname: De Simone
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D051380:GRB2 Adaptor Protein / Q000737:chemistry / Q000378:metabolism; D064267:Intrinsically Disordered Proteins / Q000737:chemistry* / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017354:Point Mutation; D011485:Protein Binding; D018909:src Homology Domains
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; D064267:Intrinsically Disordered Proteins; D050505:Mutant Proteins; D010455:Peptides
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