Structure of mouse protocadherin 15 of the stereocilia tip link in complex with LHFPL5

  1. Jingpeng Ge
  2. Johannes Elferich
  3. April Goehring
  4. Joy Zhao
  5. Peter Schuck
  6. Eric Gouaux  Is a corresponding author
  1. Oregon Health and Science University, United States
  2. National Institutes of Health, United States

Abstract

Hearing and balance involve the transduction of mechanical stimuli into electrical signals by deflection of bundles of stereocilia linked together by protocadherin 15 (PCDH15) and cadherin 23 'tip links'. PCDH15 transduces tip link tension into opening of a mechano-electrical transduction (MET) ion channel. PCDH15 also interacts with LHFPL5, a candidate subunit of the MET channel. Here we illuminate the PCDH15-LHFPL5 structure, showing how the complex is composed of PCDH15 and LHFPL5 subunit pairs related by a 2-fold axis. The extracellular cadherin domains define a mobile tether coupled to a rigid, 2-fold symmetric 'collar' proximal to the membrane bilayer. LHFPL5 forms extensive interactions with the PCDH15 transmembrane helices and stabilizes the overall PCDH15-LHFPL5 assembly. Our studies illuminate the architecture of the PCDH15-LHFPL5 complex, localize mutations associated with deafness, and shed new light on how forces in the PCDH15 tether may be transduced into the stereocilia membrane.

Data availability

The crystal structure of EC11-EL has been deposited to the Protein Data Bank under accession code 6C10. The three-dimensional cryo-EM density maps of the PCDH154EC-LHFPL5 complex and the PCDH151EC-LHFPL5 complex have been deposited to the EM database under the accession codes EMD-7327 and EMD-7328, respectively, and the coordinates for the structures have been deposited to the Protein Data Bank under the accession codes 6C13 and 6C14, respectively.

The following data sets were generated
    1. Gouaux E
    2. Elferich J
    3. Ge J
    (2018) Soluble domain of a membrane protein
    Publicly available at the RCSB Protein Data Bank (accession no: PDB 6C10).
    1. Gouaux E
    2. Elferich J
    3. Ge J
    (2018) Structure of a membrane protein complex
    Publicly available at the RCSB Protein Data Bank (accession no: PDB 6C13).
    1. Gouaux E
    2. Elferich J
    3. Ge J
    (2018) Structure of a membrane protein complex
    Publicly available at the Electron Microscopy Data Bank (accession no: EMD-7327).
    1. Gouaux E
    2. Elferich J
    3. Ge J
    (2018) cryoEM structure of membrane protein complex
    Publicly available at the RCSB Protein Data Bank (accession no: PDB 6C14).
    1. Gouaux E
    2. Elferich J
    3. Ge J
    (2018) cryoEM structure of membrane protein complex
    Publicly available at the Electron Microscopy Data Bank (accession no: EMD-7328).

Article and author information

Author details

  1. Jingpeng Ge

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6164-1221
  2. Johannes Elferich

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9911-706X
  3. April Goehring

    Vollum Institute, Oregon Health and Science University, Portland, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Joy Zhao

    Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Peter Schuck

    Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8859-6966
  6. Eric Gouaux

    Vollum Institute, Oregon Health and Science University, Portland, United States
    For correspondence
    gouauxe@ohsu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8549-2360

Funding

National Institutes of Health (NIBIB intramural research program)

  • Joy Zhao
  • Peter Schuck

Howard Hughes Medical Institute

  • Eric Gouaux

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 4,779
    views
  • 909
    downloads
  • 68
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Jingpeng Ge
  2. Johannes Elferich
  3. April Goehring
  4. Joy Zhao
  5. Peter Schuck
  6. Eric Gouaux
(2018)
Structure of mouse protocadherin 15 of the stereocilia tip link in complex with LHFPL5
eLife 7:e38770.
https://doi.org/10.7554/eLife.38770

Share this article

https://doi.org/10.7554/eLife.38770

Further reading

    1. Structural Biology and Molecular Biophysics
    Bradley P Clarke, Alexia E Angelos ... Yi Ren
    Research Article

    In eukaryotes, RNAs transcribed by RNA Pol II are modified at the 5′ end with a 7-methylguanosine (m7G) cap, which is recognized by the nuclear cap binding complex (CBC). The CBC plays multiple important roles in mRNA metabolism, including transcription, splicing, polyadenylation, and export. It promotes mRNA export through direct interaction with a key mRNA export factor, ALYREF, which in turn links the TRanscription and EXport (TREX) complex to the 5′ end of mRNA. However, the molecular mechanism for CBC-mediated recruitment of the mRNA export machinery is not well understood. Here, we present the first structure of the CBC in complex with an mRNA export factor, ALYREF. The cryo-EM structure of CBC-ALYREF reveals that the RRM domain of ALYREF makes direct contact with both the NCBP1 and NCBP2 subunits of the CBC. Comparing CBC-ALYREF with other cellular complexes containing CBC and/or ALYREF components provides insights into the coordinated events during mRNA transcription, splicing, and export.

    1. Structural Biology and Molecular Biophysics
    Yao Chi Chen, Karen Sargsyan ... Carmay Lim
    Research Article

    Experimental detection of residues critical for protein–protein interactions (PPI) is a time-consuming, costly, and labor-intensive process. Hence, high-throughput PPI-hot spot prediction methods have been developed, but they have been validated using relatively small datasets, which may compromise their predictive reliability. Here, we introduce PPI-hotspotID, a novel method for identifying PPI-hot spots using the free protein structure, and validated it on the largest collection of experimentally confirmed PPI-hot spots to date. We explored the possibility of detecting PPI-hot spots using (i) FTMap in the PPI mode, which identifies hot spots on protein–protein interfaces from the free protein structure, and (ii) the interface residues predicted by AlphaFold-Multimer. PPI-hotspotID yielded better performance than FTMap and SPOTONE, a webserver for predicting PPI-hot spots given the protein sequence. When combined with the AlphaFold-Multimer-predicted interface residues, PPI-hotspotID yielded better performance than either method alone. Furthermore, we experimentally verified several PPI-hotspotID-predicted PPI-hot spots of eukaryotic elongation factor 2. Notably, PPI-hotspotID can reveal PPI-hot spots not obvious from complex structures, including those in indirect contact with binding partners. PPI-hotspotID serves as a valuable tool for understanding PPI mechanisms and aiding drug design. It is available as a web server (https://ppihotspotid.limlab.dnsalias.org/) and open-source code (https://github.com/wrigjz/ppihotspotid/).