1. Structural Biology and Molecular Biophysics

A novel pH-dependent membrane peptide that binds to EphA2 and inhibits cell migration

  1. Daiane Santana Alves
  2. Justin M Westerfield
  3. Xiaojun Shi
  4. Vanessa P Nguyen
  5. Katherine M Stefanski
  6. Kristen R Booth
  7. Soyeon Kim
  8. Jennifer Morrell-Falvey
  9. Bing-Cheng Wang
  10. Steven M Abel
  11. Adam W Smith
  12. Francisco N Barrera  Is a corresponding author
  1. University of Tennessee, United States
  2. University of Akron, United States
  3. Case Western Reserve University, United States
https://doi.org/10.7554/eLife.36645
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  1. Daiane Santana Alves
  2. Justin M Westerfield
  3. Xiaojun Shi
  4. Vanessa P Nguyen
  5. Katherine M Stefanski
  6. Kristen R Booth
  7. Soyeon Kim
  8. Jennifer Morrell-Falvey
  9. Bing-Cheng Wang
  10. Steven M Abel
  11. Adam W Smith
  12. Francisco N Barrera
(2018)
A novel pH-dependent membrane peptide that binds to EphA2 and inhibits cell migration
eLife 7:e36645.
https://doi.org/10.7554/eLife.36645
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Abstract

Misregulation of the signaling axis formed by the receptor tyrosine kinase (RTK) EphA2 and its ligand, ephrinA1, causes aberrant cell-cell contacts that contribute to metastasis. Solid tumors are characterized by an acidic extracellular medium. We intend to take advantage of this tumor feature to design new molecules that specifically target tumors. We created a novel pH-dependent transmembrane peptide, TYPE7, by altering the sequence of the transmembrane domain of EphA2. TYPE7 is highly soluble and interacts with the surface of lipid membranes at neutral pH, while acidity triggers transmembrane insertion. TYPE7 binds to endogenous EphA2 and reduces Akt phosphorylation and cell migration as effectively as ephrinA1. Interestingly, we found large differences in juxtamembrane tyrosine phosphorylation and the extent of EphA2 clustering when compared TYPE7 with activation by ephrinA1. This work shows that it is possible to design new pH-triggered membrane peptides to activate RTK and gain insights on its activation mechanism.

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All data generated or analysed during this study are included in the manuscript and supporting files.

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Author details

  1. Daiane Santana Alves

    Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoville, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Justin M Westerfield

    Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3937-5833

  3. Xiaojun Shi

    Department of Chemistry, University of Akron, Akron, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8060-5880

  4. Vanessa P Nguyen

    Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Katherine M Stefanski

    Graduate School of Genome Science and Technology, University of Tennessee, Knoxville, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Kristen R Booth

    Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Soyeon Kim

    Department of Chemistry, University of Akron, Akron, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Jennifer Morrell-Falvey

    Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9362-7528

  9. Bing-Cheng Wang

    Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, United States
    Competing interests
    The authors declare that no competing interests exist.

  10. Steven M Abel

    Department of Chemical and Biomolecular Engineering, University of Tennessee, Knoxville, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0491-8647

  11. Adam W Smith

    Department of Chemistry, University of Akron, Akron, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5216-9017

  12. Francisco N Barrera

    Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, United States
    For correspondence
    fbarrera@utk.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5200-7891

Funding

National Institute of General Medical Sciences (R01GM120642)

  • Francisco N Barrera

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

Copyright

© 2018, Alves 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. Daiane Santana Alves
  2. Justin M Westerfield
  3. Xiaojun Shi
  4. Vanessa P Nguyen
  5. Katherine M Stefanski
  6. Kristen R Booth
  7. Soyeon Kim
  8. Jennifer Morrell-Falvey
  9. Bing-Cheng Wang
  10. Steven M Abel
  11. Adam W Smith
  12. Francisco N Barrera
(2018)
A novel pH-dependent membrane peptide that binds to EphA2 and inhibits cell migration
eLife 7:e36645.
https://doi.org/10.7554/eLife.36645

Share this article

https://doi.org/10.7554/eLife.36645

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Further reading

    1. Structural Biology and Molecular Biophysics

    Allosteric inhibition of the T cell receptor by a designed membrane ligand

    Yujie Ye, Shumpei Morita ... Francisco N Barrera
    Research Advance

    The T cell receptor (TCR) is a complex molecular machine that directs the activation of T cells, allowing the immune system to fight pathogens and cancer cells. Despite decades of investigation, the molecular mechanism of TCR activation is still controversial. One of the leading activation hypotheses is the allosteric model. This model posits that binding of pMHC at the extracellular domain triggers a dynamic change in the transmembrane (TM) domain of the TCR subunits, which leads to signaling at the cytoplasmic side. We sought to test this hypothesis by creating a TM ligand for TCR. Previously we described a method to create a soluble peptide capable of inserting into membranes and binding to the TM domain of the receptor tyrosine kinase EphA2 (Alves et al., eLife, 2018). Here, we show that the approach is generalizable to complex membrane receptors, by designing a TM ligand for TCR. We observed that the designed peptide caused a reduction of Lck phosphorylation of TCR at the CD3ζ subunit in T cells. As a result, in the presence of this peptide inhibitor of TCR (PITCR), the proximal signaling cascade downstream of TCR activation was significantly dampened. Co-localization and co-immunoprecipitation in diisobutylene maleic acid (DIBMA) native nanodiscs confirmed that PITCR was able to bind to the TCR. AlphaFold-Multimer predicted that PITCR binds to the TM region of TCR, where it interacts with the two CD3ζ subunits. Our results additionally indicate that PITCR disrupts the allosteric changes in the compactness of the TM bundle that occur upon TCR activation, lending support to the allosteric TCR activation model. The TCR inhibition achieved by PITCR might be useful to treat inflammatory and autoimmune diseases and to prevent organ transplant rejection, as in these conditions aberrant activation of TCR contributes to disease.