Analysis of the Mechanosensor Channel Functionality of TACAN

  1. Yiming Niu
  2. Xiao Tao
  3. George Vaisey
  4. Paul D B Olinares
  5. Hanan Alwaseem
  6. Brian Chait
  7. Roderick MacKinnon  Is a corresponding author
  1. Howard Hughes Medical Institute, The Rockefeller University, United States
  2. Rockefeller University, United States
  3. The Rockefeller University, United States

Abstract

Mechanosensitive ion channels mediate transmembrane ion currents activated by mechanical forces. A mechanosensitive ion channel called TACAN was recently reported. We began to study TACAN with the intent to understand how it senses mechanical forces and functions as an ion channel. Using cellular patch-recording methods we failed to identify mechanosensitive ion channel activity. Using membrane reconstitution methods we found that TACAN, at high protein concentrations, produces heterogeneous conduction levels that are not mechanosensitive and are most consistent with disruptions of the lipid bilayer. We determined the structure of TACAN using single particle cryo-EM and observe that it forms a symmetrical dimeric transmembrane protein. Each protomer contains an intracellular-facing cleft with a coenzyme-A cofactor, confirmed by mass spectrometry. The TACAN protomers are related in 3-dimensional structure to a fatty acid elongase, ELOVL. Whilst its physiological function remains unclear, we anticipate that TACAN is not a mechanosensitive ion channel.

Data availability

The B-factor sharpened 3D cryo-EM density map and atomic coordinates of wild-type TACAN have been deposited in Worldwide Protein Data Bank (wwPDB) under accession codes 7N0K and EMD-24107.The B-factor sharpened 3D cryo-EM density map and atomic coordinates of His196Ala, His197Ala mutant TACAN have been deposited in Worldwide Protein Data Bank (wwPDB) under accession codes 7N0L and EMD-24108.

The following data sets were generated

Article and author information

Author details

  1. Yiming Niu

    Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5683-1781
  2. Xiao Tao

    Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9381-7903
  3. George Vaisey

    Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Paul D B Olinares

    Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3429-6618
  5. Hanan Alwaseem

    Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Brian Chait

    Rockefeller University, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Roderick MacKinnon

    Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, United States
    For correspondence
    mackinn@mail.rockefeller.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7605-4679

Funding

National Institutes of Health (GM43949)

  • Roderick MacKinnon

National Institutes of Health (GM109824)

  • Brian Chait

National Institutes of Health (GM103314)

  • Brian Chait

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

Reviewing Editor

  1. Kenton J Swartz, National Institute of Neurological Disorders and Stroke, National Institutes of Health, United States

Publication history

  1. Received: June 11, 2021
  2. Preprint posted: June 12, 2021 (view preprint)
  3. Accepted: August 6, 2021
  4. Accepted Manuscript published: August 10, 2021 (version 1)
  5. Version of Record published: August 19, 2021 (version 2)

Copyright

© 2021, Niu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Yiming Niu
  2. Xiao Tao
  3. George Vaisey
  4. Paul D B Olinares
  5. Hanan Alwaseem
  6. Brian Chait
  7. Roderick MacKinnon
(2021)
Analysis of the Mechanosensor Channel Functionality of TACAN
eLife 10:e71188.
https://doi.org/10.7554/eLife.71188

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