A toolkit for studying cell surface shedding of diverse transmembrane receptors

  1. Amanda N Hayward
  2. Eric J Aird
  3. Wendy R Gordon  Is a corresponding author
  1. University of Minnesota, United States

Abstract

Proteolysis of transmembrane receptors is a critical cellular communication mechanism dysregulated in disease, yet decoding proteolytic regulation mechanisms of hundreds of shed receptors is hindered by difficulties controlling stimuli and unknown fates of cleavage products. Notch proteolytic regulation is a notable exception, where intercellular forces drive exposure of a cryptic protease site within a juxtamembrane proteolytic switch domain to activate transcriptional programs. We created a Synthetic Notch Assay for Proteolytic Switches (SNAPS) that exploits the modularity and unequivocal input/response of Notch proteolysis to screen surface receptors for other putative proteolytic switches. We identify several new proteolytic switches among receptors with structural homology to Notch. We demonstrate SNAPS can detect shedding in chimeras of diverse cell surface receptors, leading to new, testable hypotheses. Finally, we establish the assay can be used to measure modulation of proteolysis by potential therapeutics and offer new mechanistic insights into how DECMA-1 disrupts cell adhesion.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 1 and 3 and 4.

Article and author information

Author details

  1. Amanda N Hayward

    Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Eric J Aird

    Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4873-042X
  3. Wendy R Gordon

    Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, United States
    For correspondence
    wrgordon@umn.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7696-5560

Funding

National Institute of General Medical Sciences (R35GM119483)

  • Wendy R Gordon

Pew Charitable Trusts (Pew Biomedical Scholar)

  • Wendy R Gordon

National Cancer Institute (U54CA210190)

  • Eric J Aird

3M (Graduate student fellowship)

  • Eric J Aird

American Heart Association (Graduate student fellowship)

  • Amanda N Hayward

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

Reviewing Editor

  1. Matthew Freeman, University of Oxford, United Kingdom

Publication history

  1. Received: March 19, 2019
  2. Accepted: June 7, 2019
  3. Accepted Manuscript published: June 7, 2019 (version 1)
  4. Version of Record published: June 20, 2019 (version 2)

Copyright

© 2019, Hayward 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. Amanda N Hayward
  2. Eric J Aird
  3. Wendy R Gordon
(2019)
A toolkit for studying cell surface shedding of diverse transmembrane receptors
eLife 8:e46983.
https://doi.org/10.7554/eLife.46983

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