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

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.

Data availability

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

Article and author information

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.

Reviewing Editor

  1. Patricia Bassereau, Institut Curie, France

Version history

  1. Received: March 13, 2018
  2. Accepted: September 16, 2018
  3. Accepted Manuscript published: September 17, 2018 (version 1)
  4. Version of Record published: October 17, 2018 (version 2)
  5. Version of Record updated: August 30, 2019 (version 3)

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.

Metrics

  • 2,911
    Page views
  • 443
    Downloads
  • 30
    Citations

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

Further reading

    1. Cell Biology
    2. Structural Biology and Molecular Biophysics
    Shun Kai Yang, Shintaroh Kubo ... Khanh Huy Bui
    Research Article

    Acetylation of α-tubulin at the lysine 40 residue (αK40) by αTAT1/MEC-17 acetyltransferase modulates microtubule properties and occurs in most eukaryotic cells. Previous literatures suggest that acetylated microtubules are more stable and damage resistant. αK40 acetylation is the only known microtubule luminal post-translational modification site. The luminal location suggests that the modification tunes the lateral interaction of protofilaments inside the microtubule. In this study, we examined the effect of tubulin acetylation on the doublet microtubule (DMT) in the cilia of Tetrahymena thermophila using a combination of cryo-electron microscopy, molecular dynamics, and mass spectrometry. We found that αK40 acetylation exerts a small-scale effect on the DMT structure and stability by influencing the lateral rotational angle. In addition, comparative mass spectrometry revealed a link between αK40 acetylation and phosphorylation in cilia.

    1. Structural Biology and Molecular Biophysics
    Sebastian Jojoa-Cruz, Adrienne E Dubin ... Andrew B Ward
    Research Advance

    The dimeric two-pore OSCA/TMEM63 family has recently been identified as mechanically activated ion channels. Previously, based on the unique features of the structure of OSCA1.2, we postulated the potential involvement of several structural elements in sensing membrane tension (Jojoa-Cruz et al., 2018). Interestingly, while OSCA1, 2, and 3 clades are activated by membrane stretch in cell-attached patches (i.e. they are stretch-activated channels), they differ in their ability to transduce membrane deformation induced by a blunt probe (poking). Here, in an effort to understand the domains contributing to mechanical signal transduction, we used cryo-electron microscopy to solve the structure of Arabidopsis thaliana (At) OSCA3.1, which, unlike AtOSCA1.2, only produced stretch- but not poke-activated currents in our initial characterization (Murthy et al., 2018). Mutagenesis and electrophysiological assessment of conserved and divergent putative mechanosensitive features of OSCA1.2 reveal a selective disruption of the macroscopic currents elicited by poking without considerable effects on stretch-activated currents (SAC). Our results support the involvement of the amphipathic helix and lipid-interacting residues in the membrane fenestration in the response to poking. Our findings position these two structural elements as potential sources of functional diversity within the family.