New insights into the role of the disordered WIP N-terminal domain revealed by NMR structural characterization

Hila Elazari-Shalom, Hadassa Shaked, Santiago Esteban-Martin, Xavier Salvatella, Mira Barda-Saad, Jordan H. Chill

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

WASp-interacting protein (WIP) is an intrinsically disordered 503-residue polypeptide with a key role in actin polymerization in activated T cells. Its interaction with actin is mediated by a pair of conserved actin binding motifs (ABMs) at the WIP N-terminus, a domain that has not been investigated in its unbound form. Here we use NMR to investigate the biophysical behavior of the N-terminal ABM in WIP using protonless 13C-detected spectroscopy. Secondary chemical shifts, residual dipolar couplings and temperature effects identify residual structure throughout the ABM, which exhibits transient helical and β-strand character for residues 30-42 and 44-62, respectively. These observed structural propensities echo the structure observed in the actin-bound state of the ABM. Furthermore, residues preceding the canonical ABM (17-25) and conserved among WIP-related proteins exhibit transient β-strand character, suggesting that the WIPN interaction epitope extends towards the N-terminal polyproline motif. This suggests a possible role for this region in mediating the WIP interaction with polyproline binders such as profilin. In revealing these features of the WIP ABM this study demonstrates the unique ability of NMR in characterizing unstructured domains and provides necessary information for further investigation of WIP-mediated protein-protein interactions.

Original languageEnglish
Pages (from-to)700-714
Number of pages15
JournalFEBS Journal
Volume282
Issue number4
DOIs
StatePublished - Feb 2015

Bibliographical note

Publisher Copyright:
© 2014 FEBS.

Keywords

  • C-detected spectroscopy
  • NMR
  • Wiskott-Aldrich syndrome
  • actin binding motif
  • intrinsically disordered proteins
  • structural biology

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