Biophysical basis of the binding of WWOX tumor suppressor to WBP1 and WBP2 adaptors

Caleb B. McDonald, Laura Buffa, Tomer Bar-Mag, Zaidoun Salah, Vikas Bhat, David C. Mikles, Brian J. Deegan, Kenneth L. Seldeen, Arun Malhotra, Marius Sudol, Rami I. Aqeilan, Zafar Nawaz, Amjad Farooq

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The WW-containing oxidoreductase (WWOX) tumor suppressor participates in a diverse array of cellular activities by virtue of its ability to recognize WW-binding protein 1 (WBP1) and WW-binding protein 2 (WBP2) signaling adaptors among a wide variety of other ligands. Herein, using a multitude of biophysical techniques, we provide evidence that while the WW1 domain of WWOX binds to PPXY motifs within WBP1 and WBP2 in a physiologically relevant manner, the WW2 domain exhibits no affinity toward any of these PPXY motifs. Importantly, our data suggest that while R25/W44 residues located within the binding pocket of a triple-stranded β-fold of WW1 domain are critical for the recognition of PPXY ligands, they are replaced by the chemically distinct E66/Y85 duo at structurally equivalent positions within the WW2 domain, thereby accounting for its failure to bind PPXY ligands. Predictably, not only does the introduction of E66R/Y85W double substitution within the WW2 domain result in gain of function but the resulting engineered domain, hereinafter referred to as WW2-RW, also appears to be a much stronger binding partner of WBP1 and WBP2 than the wild-type WW1 domain. We also show that while the WW1 domain is structurally disordered and folds upon ligand binding, the WW2 domain not only adopts a fully structured conformation but also aids stabilization and ligand binding to WW1 domain. This salient observation implies that the WW2 domain likely serves as a chaperone to augment the physiological function of WW1 domain within WWOX. Collectively, our study lays the groundwork for understanding the molecular basis of a key protein-protein interaction pertinent to human health and disease.

Original languageEnglish
Pages (from-to)58-74
Number of pages17
JournalJournal of Molecular Biology
Volume422
Issue number1
DOIs
StatePublished - Sep 7 2012

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Carrier Proteins
Oxidoreductases
Neoplasms
Ligands
Proteins
Observation
Health
PPXY ligand

Keywords

  • WW domain chaperone
  • WW domain engineering
  • WW tandem domains
  • WW-ligand thermodynamics

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Biophysical basis of the binding of WWOX tumor suppressor to WBP1 and WBP2 adaptors. / McDonald, Caleb B.; Buffa, Laura; Bar-Mag, Tomer; Salah, Zaidoun; Bhat, Vikas; Mikles, David C.; Deegan, Brian J.; Seldeen, Kenneth L.; Malhotra, Arun; Sudol, Marius; Aqeilan, Rami I.; Nawaz, Zafar; Farooq, Amjad.

In: Journal of Molecular Biology, Vol. 422, No. 1, 07.09.2012, p. 58-74.

Research output: Contribution to journalArticle

McDonald, CB, Buffa, L, Bar-Mag, T, Salah, Z, Bhat, V, Mikles, DC, Deegan, BJ, Seldeen, KL, Malhotra, A, Sudol, M, Aqeilan, RI, Nawaz, Z & Farooq, A 2012, 'Biophysical basis of the binding of WWOX tumor suppressor to WBP1 and WBP2 adaptors', Journal of Molecular Biology, vol. 422, no. 1, pp. 58-74. https://doi.org/10.1016/j.jmb.2012.05.015
McDonald, Caleb B. ; Buffa, Laura ; Bar-Mag, Tomer ; Salah, Zaidoun ; Bhat, Vikas ; Mikles, David C. ; Deegan, Brian J. ; Seldeen, Kenneth L. ; Malhotra, Arun ; Sudol, Marius ; Aqeilan, Rami I. ; Nawaz, Zafar ; Farooq, Amjad. / Biophysical basis of the binding of WWOX tumor suppressor to WBP1 and WBP2 adaptors. In: Journal of Molecular Biology. 2012 ; Vol. 422, No. 1. pp. 58-74.
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AU - Bhat, Vikas

AU - Mikles, David C.

AU - Deegan, Brian J.

AU - Seldeen, Kenneth L.

AU - Malhotra, Arun

AU - Sudol, Marius

AU - Aqeilan, Rami I.

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