Polyunsaturated fatty acids inhibit a pentameric ligand-gated ion channel through one of two binding sites

  1. Noah M Dietzen
  2. Mark J Arcario
  3. Lawrence J Chen
  4. John T Petroff II
  5. K Trent Moreland
  6. Kathiresan Krishnan
  7. Grace Brannigan
  8. Douglas F Covey
  9. Wayland WL Cheng  Is a corresponding author
  1. Washington University in St. Louis, United States
  2. Rutgers University, United States

Abstract

Polyunsaturated fatty acids (PUFAs) inhibit pentameric ligand-gated ion channels (pLGICs) but the mechanism of inhibition is not well understood. The PUFA, docosahexaenoic acid (DHA), inhibits agonist responses of the pLGIC, ELIC, more effectively than palmitic acid, similar to the effects observed in the GABAA receptor and nicotinic acetylcholine receptor. Using photo-affinity labeling and coarse-grained molecular dynamics simulations, we identified two fatty acid binding sites in the outer transmembrane domain (TMD) of ELIC. Fatty acid binding to the photolabeled sites is selective for DHA over palmitic acid, and specific for an agonist-bound state. Hexadecyl-methanethiosulfonate modification of one of the two fatty acid binding sites in the outer TMD recapitulates the inhibitory effect of PUFAs in ELIC. The results demonstrate that DHA selectively binds to multiple sites in the outer TMD of ELIC, but that state-dependent binding to a single intrasubunit site mediates DHA inhibition of ELIC.

Data availability

Figure 4- source data 1 contains the numerical data used to generate Figure 4A and 4B. Figure 4- source data 2 contains the numerical data used to generate Figure 4C and Figure 4- figure supplement 5. Figure 4- source data 3 contains the statistical analysis (linear mixed effects model) for Figure 4- figure supplement 5.

Article and author information

Author details

  1. Noah M Dietzen

    Department of Anesthesiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Mark J Arcario

    Department of Anesthesiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5017-1519
  3. Lawrence J Chen

    Department of Anesthesiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. John T Petroff II

    Department of Anesthesiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1323-0273
  5. K Trent Moreland

    Department of Anesthesiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Kathiresan Krishnan

    Department of Developmental Biology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Grace Brannigan

    Center for the Computational and Integrative Biology, Rutgers University, Camden, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Douglas F Covey

    Department of Developmental Biology, Washington University in St. Louis, St Louis, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Wayland WL Cheng

    Department of Anesthesiology, Washington University in St. Louis, St Louis, United States
    For correspondence
    wayland.cheng@wustl.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9529-9820

Funding

National Institutes of Health (R35GM137957)

  • Wayland WL Cheng

National Institutes of Health (F32GM139351)

  • John T Petroff II

National Institutes of Health (R01HL067773)

  • Douglas F Covey

National Institutes of Health (R01GM108799)

  • Douglas F Covey

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

Reviewing Editor

  1. Marcel P Goldschen-Ohm, University of Texas at Austin, United States

Publication history

  1. Received: September 29, 2021
  2. Preprint posted: October 8, 2021 (view preprint)
  3. Accepted: December 31, 2021
  4. Accepted Manuscript published: January 4, 2022 (version 1)
  5. Accepted Manuscript updated: January 7, 2022 (version 2)
  6. Version of Record published: January 24, 2022 (version 3)

Copyright

© 2022, Dietzen 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.

Metrics

  • 1,127
    Page views
  • 221
    Downloads
  • 3
    Citations

Article citation count generated by polling the highest count across the following sources: PubMed Central, Crossref, Scopus.

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. Noah M Dietzen
  2. Mark J Arcario
  3. Lawrence J Chen
  4. John T Petroff II
  5. K Trent Moreland
  6. Kathiresan Krishnan
  7. Grace Brannigan
  8. Douglas F Covey
  9. Wayland WL Cheng
(2022)
Polyunsaturated fatty acids inhibit a pentameric ligand-gated ion channel through one of two binding sites
eLife 11:e74306.
https://doi.org/10.7554/eLife.74306
  1. Further reading

Further reading

    1. Biochemistry and Chemical Biology
    2. Neuroscience
    Jinli Geng, Yingjun Tang ... Xiaodong Liu
    Research Article Updated

    Dynamic Ca2+ signals reflect acute changes in membrane excitability, and also mediate signaling cascades in chronic processes. In both cases, chronic Ca2+ imaging is often desired, but challenged by the cytotoxicity intrinsic to calmodulin (CaM)-based GCaMP, a series of genetically-encoded Ca2+ indicators that have been widely applied. Here, we demonstrate the performance of GCaMP-X in chronic Ca2+ imaging of cortical neurons, where GCaMP-X by design is to eliminate the unwanted interactions between the conventional GCaMP and endogenous (apo)CaM-binding proteins. By expressing in adult mice at high levels over an extended time frame, GCaMP-X showed less damage and improved performance in two-photon imaging of sensory (whisker-deflection) responses or spontaneous Ca2+ fluctuations, in comparison with GCaMP. Chronic Ca2+ imaging of one month or longer was conducted for cultured cortical neurons expressing GCaMP-X, unveiling that spontaneous/local Ca2+ transients progressively developed into autonomous/global Ca2+ oscillations. Along with the morphological indices of neurite length and soma size, the major metrics of oscillatory Ca2+, including rate, amplitude and synchrony were also examined. Dysregulations of both neuritogenesis and Ca2+ oscillations became discernible around 2–3 weeks after virus injection or drug induction to express GCaMP in newborn or mature neurons, which were exacerbated by stronger or prolonged expression of GCaMP. In contrast, neurons expressing GCaMP-X were significantly less damaged or perturbed, altogether highlighting the unique importance of oscillatory Ca2+ to neural development and neuronal health. In summary, GCaMP-X provides a viable solution for Ca2+ imaging applications involving long-time and/or high-level expression of Ca2+ probes.

    1. Biochemistry and Chemical Biology
    2. Chromosomes and Gene Expression
    Radhika A Varier, Theodora Sideri ... Folkert Jacobus van Werven
    Research Article

    N6-methyladenosine (m6A) RNA modification impacts mRNA fate primarily via reader proteins, which dictate processes in development, stress, and disease. Yet little is known about m6A function in Saccharomyces cerevisiae, which occurs solely during early meiosis. Here we perform a multifaceted analysis of the m6A reader protein Pho92/Mrb1. Cross-linking immunoprecipitation analysis reveals that Pho92 associates with the 3’end of meiotic mRNAs in both an m6A-dependent and independent manner. Within cells, Pho92 transitions from the nucleus to the cytoplasm, and associates with translating ribosomes. In the nucleus Pho92 associates with target loci through its interaction with transcriptional elongator Paf1C. Functionally, we show that Pho92 promotes and links protein synthesis to mRNA decay. As such, the Pho92-mediated m6A-mRNA decay is contingent on active translation and the CCR4-NOT complex. We propose that the m6A reader Pho92 is loaded co-transcriptionally to facilitate protein synthesis and subsequent decay of m6A modified transcripts, and thereby promotes meiosis.