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Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX
Class I WW domains are present in many proteins of various functions and mediate protein interactions by binding to short linear PPxY motifs. Tandem WW domains often bind peptides with multiple PPxY motifs, but the interplay of WW–peptide interactions is not always intuitive. The WW domain–containin...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9293652/ https://www.ncbi.nlm.nih.gov/pubmed/35716775 http://dx.doi.org/10.1016/j.jbc.2022.102145 |
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author | Rotem-Bamberger, Shahar Fahoum, Jamal Keinan-Adamsky, Keren Tsaban, Tomer Avraham, Orly Shalev, Deborah E. Chill, Jordan H. Schueler-Furman, Ora |
author_facet | Rotem-Bamberger, Shahar Fahoum, Jamal Keinan-Adamsky, Keren Tsaban, Tomer Avraham, Orly Shalev, Deborah E. Chill, Jordan H. Schueler-Furman, Ora |
author_sort | Rotem-Bamberger, Shahar |
collection | PubMed |
description | Class I WW domains are present in many proteins of various functions and mediate protein interactions by binding to short linear PPxY motifs. Tandem WW domains often bind peptides with multiple PPxY motifs, but the interplay of WW–peptide interactions is not always intuitive. The WW domain–containing oxidoreductase (WWOX) harbors two WW domains: an unstable WW1 capable of PPxY binding and stable WW2 that cannot bind PPxY. The WW2 domain has been suggested to act as a WW1 domain chaperone, but the underlying mechanism of its chaperone activity remains to be revealed. Here, we combined NMR, isothermal calorimetry, and structural modeling to elucidate the roles of both WW domains in WWOX binding to its PPxY-containing substrate ErbB4. Using NMR, we identified an interaction surface between these two domains that supports a WWOX conformation compatible with peptide substrate binding. Isothermal calorimetry and NMR measurements also indicated that while binding affinity to a single PPxY motif is marginally increased in the presence of WW2, affinity to a dual-motif peptide increases 10-fold. Furthermore, we found WW2 can directly bind double-motif peptides using its canonical binding site. Finally, differential binding of peptides in mutagenesis experiments was consistent with a parallel N- to C-terminal PPxY tandem motif orientation in binding to the WW1–WW2 tandem domain, validating structural models of the interaction. Taken together, our results reveal the complex nature of tandem WW-domain organization and substrate binding, highlighting the contribution of WWOX WW2 to both protein stability and target binding. |
format | Online Article Text |
id | pubmed-9293652 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-92936522022-07-20 Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX Rotem-Bamberger, Shahar Fahoum, Jamal Keinan-Adamsky, Keren Tsaban, Tomer Avraham, Orly Shalev, Deborah E. Chill, Jordan H. Schueler-Furman, Ora J Biol Chem Research Article Class I WW domains are present in many proteins of various functions and mediate protein interactions by binding to short linear PPxY motifs. Tandem WW domains often bind peptides with multiple PPxY motifs, but the interplay of WW–peptide interactions is not always intuitive. The WW domain–containing oxidoreductase (WWOX) harbors two WW domains: an unstable WW1 capable of PPxY binding and stable WW2 that cannot bind PPxY. The WW2 domain has been suggested to act as a WW1 domain chaperone, but the underlying mechanism of its chaperone activity remains to be revealed. Here, we combined NMR, isothermal calorimetry, and structural modeling to elucidate the roles of both WW domains in WWOX binding to its PPxY-containing substrate ErbB4. Using NMR, we identified an interaction surface between these two domains that supports a WWOX conformation compatible with peptide substrate binding. Isothermal calorimetry and NMR measurements also indicated that while binding affinity to a single PPxY motif is marginally increased in the presence of WW2, affinity to a dual-motif peptide increases 10-fold. Furthermore, we found WW2 can directly bind double-motif peptides using its canonical binding site. Finally, differential binding of peptides in mutagenesis experiments was consistent with a parallel N- to C-terminal PPxY tandem motif orientation in binding to the WW1–WW2 tandem domain, validating structural models of the interaction. Taken together, our results reveal the complex nature of tandem WW-domain organization and substrate binding, highlighting the contribution of WWOX WW2 to both protein stability and target binding. American Society for Biochemistry and Molecular Biology 2022-06-16 /pmc/articles/PMC9293652/ /pubmed/35716775 http://dx.doi.org/10.1016/j.jbc.2022.102145 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Rotem-Bamberger, Shahar Fahoum, Jamal Keinan-Adamsky, Keren Tsaban, Tomer Avraham, Orly Shalev, Deborah E. Chill, Jordan H. Schueler-Furman, Ora Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX |
title | Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX |
title_full | Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX |
title_fullStr | Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX |
title_full_unstemmed | Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX |
title_short | Structural insights into the role of the WW2 domain on tandem WW–PPxY motif interactions of oxidoreductase WWOX |
title_sort | structural insights into the role of the ww2 domain on tandem ww–ppxy motif interactions of oxidoreductase wwox |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9293652/ https://www.ncbi.nlm.nih.gov/pubmed/35716775 http://dx.doi.org/10.1016/j.jbc.2022.102145 |
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