Recognition of lysine-rich peptide ligands by murine cortactin SH3 domain: CD, ITC, and NMR studies

Biopolymers
Chiara RubiniArianna Donella-Deana

Abstract

Cortactin is a ubiquitous actin-binding protein that regulates various aspects of cell dynamics and is implicated in the pathogenesis of human neoplasia. The sequence of cortactin contains a number of signaling motifs and an SH3 domain at the C-terminus, which mediates the interaction of the protein with several partners, including Shank2. A recombinant protein, comprising the murine cortactin SH3 domain fused to GST (GST-SH3(m-cort)), was prepared and used to assess the domain-binding affinity of potential peptide-ligands reproducing the proline-rich regions of human HPK1 and Shank2 proteins. The key residues involved in the SH3(m-cort) domain recognition were identified by three different approaches: non-immobilized ligand interaction assay by circular dichroism, isothermal titration calorimetry, and nuclear magnetic resonance. Our results show that the classical PxxPxK class II binding motif is not sufficient to mediate the interaction with GST-SH3(m-cort), an event that depends on the presence of additional basic residues located at either the N- or the C-terminus of the PxxPxK motif. Especially effective in promoting the peptide binding is a Lys residue at the -5 position, a determinant present in both P2 (HPK1 394-403) an...Continue Reading

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Citations

Jan 29, 2013·Small GTPases·B Daniel Lam, Peter L Hordijk
Jun 23, 2016·Biophysical Journal·Veer S BhattJae-Hyun Cho
Feb 3, 2011·Amino Acids·Giuliano SiligardiArianna Donella-Deana
Jan 4, 2012·Journal of Molecular Recognition : JMR·Rajesh GhaiBrett M Collins
Jan 19, 2020·Biophysics Reviews·Himal Kanti Ganguly, Gautam Basu

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