Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Solution structure of a Nedd4 WW domain–ENaC peptide complex

Nature Structural Biology volume 8, pages 407–412 (2001)Cite this article

Abstract

Nedd4 is a ubiquitin protein ligase composed of a C2 domain, three (or four) WW domains and a ubiquitin ligase Hect domain. Nedd4 was demonstrated to bind the epithelial sodium channel (αβγENaC), by association of its WW domains with PY motifs (XPPXY) present in each ENaC subunit, and to regulate the cell surface stability of the channel. The PY motif of βENaC is deleted or mutated in Liddle syndrome, a hereditary form of hypertension caused by elevated ENaC activity. Here we report the solution structure of the third WW domain of Nedd4 complexed to the PY motif-containing region of βENaC (TLPIPGTPPPNYDSL, referred to as βP2). A polyproline type II helical conformation is adopted by the PPPN sequence. Unexpectedly, the C-terminal sequence YDSL forms a helical turn and both the tyrosine and the C-terminal leucine contact the WW domain. This is unlike other proline-rich peptides complexed to WW domains, which bind in an extended conformation and lack molecular interactions with residues C-terminal to the tyrosine or the structurally equivalent residue in non-PY motif WW domain targets. The Nedd4 WW domain–ENaC βP2 peptide structure expands our understanding of the mechanisms involved in WW domain–ligand recognition and the molecular basis of Liddle syndrome.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Solution structure of the rNedd4 WWIII–rENaC βP2 peptide complex.
Figure 2: The rNedd4 WWIII–rENaC βP2 peptide binding interface.
Figure 3: Surface representations of WW domain-ligand complexes.

Similar content being viewed by others

Accession codes

Accessions

Protein Data Bank

References

  1. Kumar, S., Tomooka, Y. & Noda, M. Biochem. Biophys. Res. Commun. 185, 1155–1161 (1992).

    Article  CAS  Google Scholar 

  2. Bork, P. & Sudol, M. Trends Biochem. Sci. 19, 531–533 (1994).

    Article  CAS  Google Scholar 

  3. Chen, H.I. & Sudol, M. Proc. Natl. Acad. Sci. USA 92, 7819–7823 (1995).

    Article  CAS  Google Scholar 

  4. Bedford, M.T., Chan, D.C. & Leder, P. EMBO J. 16, 2376–2383 (1997).

    Article  CAS  Google Scholar 

  5. Bedford, M.T., Sarbassova, D., Xu, J., Leder, P. & Yaffe, M.B. J. Biol. Chem. 275, 10359–10369 (2000).

    Article  CAS  Google Scholar 

  6. Bedford, M.T., Reed, R. & Leder, P. Proc. Natl. Acad. Sci. USA 95, 10602–10607 (1998).

    Article  CAS  Google Scholar 

  7. Verdecia, M.A., Bowman, M.E., Lu, K.P., Hunter, T. & Noel, J.P. Nature Struct. Biol. 7, 639–643 (2000).

    Article  CAS  Google Scholar 

  8. Canessa, C.M. et al. Nature 367, 463–467 (1994).

    Article  CAS  Google Scholar 

  9. Garty, H. & Palmer, L.G. Physiol. Rev. 77, 359–396 (1997).

    Article  CAS  Google Scholar 

  10. Shimkets, R.A. et al. Cell 79, 407–414 (1994).

    Article  CAS  Google Scholar 

  11. Staub, O. et al. EMBO J. 15, 2371–2380 (1996).

    Article  CAS  Google Scholar 

  12. Schild, L. et al. EMBO J. 15, 2381–2387 (1996).

    Article  CAS  Google Scholar 

  13. Hansson, J.H. et al. Nature Genet. 11, 76–82 (1995).

    Article  CAS  Google Scholar 

  14. Hansson, J.H. et al. Proc. Natl. Acad. Sci. USA 92, 11495–11499 (1995).

    Article  CAS  Google Scholar 

  15. Tamura, H. et al. J. Clin. Invest. 97, 1780–1784 (1996).

    Article  CAS  Google Scholar 

  16. Inoue, J. et al. J. Clin. Endocrinol. Metab. 83, 2210–2213 (1998).

    CAS  PubMed  Google Scholar 

  17. Staub, O. et al. EMBO J. 16, 6325–6336 (1997).

    Article  CAS  Google Scholar 

  18. Abriel, H. et al. J. Clin. Invest. 103, 667–673 (1999).

    Article  CAS  Google Scholar 

  19. Pirozzi, G. et al. J. Biol. Chem. 272, 14611–14616 (1997).

    Article  CAS  Google Scholar 

  20. Snyder, P.M., Cheng, C., Prince, L.S., Rogers, J.C. & Welsh, M.J. J. Biol. Chem. 273, 681–684 (1998).

    Article  CAS  Google Scholar 

  21. Kanelis, V., Farrow, N.A., Kay, L.E., Rotin, D. & Forman-Kay, J.D. Biochem. Cell Biol. 76, 341–350 (1998).

    Article  CAS  Google Scholar 

  22. Kamynina, E., Debonneville, C., Bens, M., Vandewalle, A. & Staub, O. Faseb J. 15, 204–214 (2001).

    Article  CAS  Google Scholar 

  23. Macias, M.J. et al. Nature 382, 646–649 (1996).

    Article  CAS  Google Scholar 

  24. Macias, M.J., Gervais, V., Civera, C. & Oschkinat, H. Nature Struct. Biol. 7, 375–379 (2000).

    Article  CAS  Google Scholar 

  25. Huang, X. et al. Nature Struct. Biol. 7, 634–638 (2000).

    Article  CAS  Google Scholar 

  26. Zarrinpar, A. & Lim, W.A. Nature Struct. Biol. 7, 611–613 (2000).

    Article  CAS  Google Scholar 

  27. Snyder, P.M. et al. Cell 83, 969–978 (1995).

    Article  CAS  Google Scholar 

  28. Boll, W. et al. EMBO J. 15, 5789–5795 (1996).

    Article  CAS  Google Scholar 

  29. Shimkets, R.A., Lifton, R.P. & Canessa, C.M. J. Biol. Chem. 272, 25537–25541 (1997).

    Article  CAS  Google Scholar 

  30. Shimkets, R.A., Lifton, R. & Canessa, C.M. Proc. Natl. Acad. Sci. USA 95, 3301–3305 (1998).

    Article  CAS  Google Scholar 

  31. Viguera, A.R., Arrondo, J.L., Musacchio, A., Saraste, M. & Serrano, L. Biochemistry 33, 10925–10933 (1994).

    Article  CAS  Google Scholar 

  32. Delaglio, F. et al. J. Biomol. NMR 6, 277–293 (1995).

    Article  CAS  Google Scholar 

  33. Johnson, B.A. & Blevins, R.A. J. Biomol. NMR 4, 603–614 (1994).

    Article  CAS  Google Scholar 

  34. Kay, L.E. Prog. Biophys. Mol. Biol. 63, 277–299 (1995).

    Article  CAS  Google Scholar 

  35. Sattler, M., Schleucher, J. & Griesinger, C. Prog. NMR Spect. 34, 93–158 (1999).

    Article  CAS  Google Scholar 

  36. Kanelis, V. et al. J. Biomol. NMR 16, 253–259 (2000).

    Article  CAS  Google Scholar 

  37. Yamazaki, T., Forman-Kay, J.D. & Kay, L.E. J. Am. Chem. Soc. 115, 11054–11055 (1993).

    Article  CAS  Google Scholar 

  38. Pelton, J.G., Torchia, D.A., Meadow, N.D. & Roseman, S. Protein Sci. 2, 543–558 (1993).

    Article  CAS  Google Scholar 

  39. Pascal, S.M., Muhandiram, D.R., Yamazaki, T., Forman-Kay, J.D. & Kay, L.E. J. Magn. Reson. 103, 197–201 (1994).

    Article  CAS  Google Scholar 

  40. Bax, A. et al. Methods Enzymol. 239, 79–105 (1994).

    Article  CAS  Google Scholar 

  41. Grzesiek, S., Kuboniwa, H., Hinck, A.P. & Bax, A. J. Am. Chem. Soc. 117 5312–5315 (1995).

    Article  CAS  Google Scholar 

  42. Bax, A., Max, D. & Zax, D. J. Am. Chem. Soc 114, 6923–6925 (1992).

    Article  CAS  Google Scholar 

  43. Zwahlen, C. et al. J. Am. Chem. Soc. 119, 6711–6721 (1997).

    Article  CAS  Google Scholar 

  44. Cordier, F. & Grzesiek, S. J. Am. Chem. Soc. 121, 1601–1602 (1999).

    Article  CAS  Google Scholar 

  45. Vuister, G.W. & Bax, A. J. Am. Chem. Soc. 115, 7772–7777 (1993).

    Article  CAS  Google Scholar 

  46. Yang, D., Gardner, K.H. & Kay, L.E. J. Biomol. NMR 11 213–220 (1998).

    Article  CAS  Google Scholar 

  47. Yang, D. & Kay, L.E. J. Am. Chem. Soc. 120, 9880–9887 (1998).

    Article  CAS  Google Scholar 

  48. Brünger, A.T. et al. Acta Crystallogr. D 54, 905–921 (1998).

    Article  Google Scholar 

  49. Garrett, D.S. et al. J. Magn. Reson. 104, 99–103 (1994).

    Article  CAS  Google Scholar 

  50. Nilges, M. & O'Donoghue, S. Prog. NMR Spect. 32, 107–139 (1998).

    Article  CAS  Google Scholar 

  51. Cornilescu, G., Delaglio, F. & Bax, A. J. Biomol. NMR 13, 289–302 (1999).

    Article  CAS  Google Scholar 

  52. Seavey, B.R., Farr, E.A., Westler, W.M. & Markley, J.L. J. Biomol. NMR 1, 217–236 (1991).

    Article  CAS  Google Scholar 

  53. Holm, L. & Sander, C. Nucleic Acids Res. 26, 316–319 (1998).

    Article  CAS  Google Scholar 

  54. Koradi, R., Billeter, M. & Wuthrich, K. J. Mol. Graph. 14, 29–32 (1996).

    Article  Google Scholar 

  55. Laskowski, R.A., Rullman, J.A., MacArthur, M.W., Kaptein, R. & Thornton, J.M. J. Biomol. NMR 8, 477–486 (1996).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We acknowledge M. Nilges for modifications to ARIA and many useful discussions. We thank R. Muhandiram, D. Yang and L.E. Kay for assistance with NMR data collection. Structures were calculated at the Ontario Centre for Genomic Computing. We thank J.P. Noel, M.J. Eck and O. Staub for preprints prior to publication. V.K. is a recipient of an Ontario Graduate Fellowship. This work was supported by grants to D.R. from the Medical Research Council of Canada (MRC) / Canadian Institute for Health Research (CIHR) and Human Frontier Science Program, and to J.D.F.–K. from the National Cancer Institute of Canada (NCIC).

Author information

Authors and Affiliations

  1. Programmes in Structural Biology and Biochemistry, Hospital for Sick Children, 555 University Ave, Toronto, M5G 1X8, Ontario, Canada

    Voula Kanelis & Julie D. Forman-Kay

  2. Cell Biology, Hospital for Sick Children, 555 University Ave, Toronto, M5G 1X8, Ontario, Canada

    Voula Kanelis & Daniela Rotin

  3. Department of Biochemistry, University of Toronto, Toronto, M5S 1A8, Ontario, Canada

    Voula Kanelis, Daniela Rotin & Julie D. Forman-Kay

Authors
  1. Voula Kanelis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniela Rotin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Julie D. Forman-Kay
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Julie D. Forman-Kay.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kanelis, V., Rotin, D. & Forman-Kay, J. Solution structure of a Nedd4 WW domain–ENaC peptide complex. Nat Struct Mol Biol 8, 407–412 (2001). https://doi.org/10.1038/87562

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/87562

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing