The molecular appearance of native TRPM7 channel complexes identified by high-resolution proteomics

  1. Astrid Kollewe
  2. Vladimir Chubanov
  3. Fong Tsuen Tseung
  4. Leonor Correia
  5. Eva Schmidt
  6. Anna Rössig
  7. Susanna Zierler
  8. Alexander Haupt
  9. Catrin Swantje Müller
  10. Wolfgang Bildl
  11. Uwe Schulte
  12. Annette Nicke  Is a corresponding author
  13. Bernd Fakler  Is a corresponding author
  14. Thomas Gudermann  Is a corresponding author
  1. University of Freiburg, Germany
  2. Ludwig Maximilian University of Munich, Germany
  3. University Linz, Austria
  4. Max Planck Institute for Experimental Medicine, Germany
  5. LMU Munich, Germany

Abstract

The transient receptor potential melastatin-subfamily member 7 (TRPM7) is a ubiquitously expressed membrane protein consisting of ion channel and protein kinase domains. TRPM7 plays a fundamental role in the cellular uptake of divalent cations such as Zn2+, Mg2+ and Ca2+, and thus shapes cellular excitability, plasticity and metabolic activity. The molecular appearance and operation of TRPM7 channel complexes in native tissues have remained unresolved. Here, we investigated the subunit composition of endogenous TRPM7 channels in rodent brain by multi-epitope affinity purification and high-resolution quantitative MS analysis. We found that native TRPM7 channels are high molecular-weight multi-protein complexes that contain the putative metal transporter proteins CNNM1-4 and a small G-protein ARL15. Heterologous reconstitution experiments confirmed the formation of TRPM7/CNNM/ARL15 ternary complexes and indicated that complex formation effectively and specifically impacts TRPM7 activity. These results open up new avenues towards a mechanistic understanding of the cellular regulation and function of TRPM7 channels.

Data availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD025279 and DOI: 10.6019/PXD025279.

Article and author information

Author details

  1. Astrid Kollewe

    Institute of Physiology, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Vladimir Chubanov

    Walther-Straub Institute of Pharmacology and Toxicology, Ludwig Maximilian University of Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6042-4193
  3. Fong Tsuen Tseung

    Walther-Straub Institute of Pharmacology and Toxicology, Ludwig Maximilian University of Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Leonor Correia

    Walther-Straub Institute of Pharmacology and Toxicology, Ludwig Maximilian University of Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Eva Schmidt

    Walther-Straub Institute of Pharmacology and Toxicology, Ludwig Maximilian University of Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Anna Rössig

    Walther-Straub Institute of Pharmacology and Toxicology, Ludwig Maximilian University of Munich, Munich, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Susanna Zierler

    University Linz, Linz, Austria
    Competing interests
    The authors declare that no competing interests exist.
  8. Alexander Haupt

    Institute of Physiology, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5647-5724
  9. Catrin Swantje Müller

    Institute of Physiology, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Wolfgang Bildl

    Institute of Physiology, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  11. Uwe Schulte

    Institute of Physiology, University of Freiburg, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  12. Annette Nicke

    Department of Molecular Biology of Neuronal Signals, Max Planck Institute for Experimental Medicine, Göttingen, Germany
    For correspondence
    annette.nicke@lrz.uni-muenchen.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6798-505X
  13. Bernd Fakler

    Institute of Physiology, University of Freiburg, Freiburg, Germany
    For correspondence
    bernd.fakler@physiologie.uni-freiburg.de
    Competing interests
    The authors declare that no competing interests exist.
  14. Thomas Gudermann

    Walther Straub Institute of Pharmacology and Toxicology, LMU Munich, Munich, Germany
    For correspondence
    Thomas.Gudermann@lrz.uni-muenchen.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0323-7965

Funding

Deutsche Forschungsgemeinschaft (TRR 152 P15)

  • Vladimir Chubanov

Deutsche Forschungsgemeinschaft (TRR 152 P15)

  • Thomas Gudermann

Deutsche Forschungsgemeinschaft (TRR 152 P02)

  • Bernd Fakler

Deutsche Forschungsgemeinschaft (TRR 152 P02)

  • Uwe Schulte

Deutsche Forschungsgemeinschaft (SFB 1328 P15)

  • Annette Nicke

Deutsche Forschungsgemeinschaft (SFB 1381)

  • Bernd Fakler

Deutsche Forschungsgemeinschaft (Research Training Group 2338)

  • Thomas Gudermann

Deutsche Forschungsgemeinschaft (Research Training Group 2338)

  • Annette Nicke

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

Reviewing Editor

  1. László Csanády, Semmelweis University, Hungary

Version history

  1. Received: March 18, 2021
  2. Preprint posted: July 9, 2021 (view preprint)
  3. Accepted: November 8, 2021
  4. Accepted Manuscript published: November 12, 2021 (version 1)
  5. Version of Record published: November 25, 2021 (version 2)

Copyright

© 2021, Kollewe 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. Astrid Kollewe
  2. Vladimir Chubanov
  3. Fong Tsuen Tseung
  4. Leonor Correia
  5. Eva Schmidt
  6. Anna Rössig
  7. Susanna Zierler
  8. Alexander Haupt
  9. Catrin Swantje Müller
  10. Wolfgang Bildl
  11. Uwe Schulte
  12. Annette Nicke
  13. Bernd Fakler
  14. Thomas Gudermann
(2021)
The molecular appearance of native TRPM7 channel complexes identified by high-resolution proteomics
eLife 10:e68544.
https://doi.org/10.7554/eLife.68544

Share this article

https://doi.org/10.7554/eLife.68544

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