A toolkit for studying cell surface shedding of diverse transmembrane receptors

Amanda N. Hayward, Eric J. Aird, Wendy R Gordon

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Article numbere46983
JournaleLife
Volume8
DOIs
StatePublished - Jun 1 2019

Fingerprint

Proteolysis
Switches
Assays
Cell Surface Receptors
Cell Adhesion
Peptide Hydrolases
Cellular radio systems
Cell adhesion
Decoding
Modulation
Therapeutics

Cite this

A toolkit for studying cell surface shedding of diverse transmembrane receptors. / Hayward, Amanda N.; Aird, Eric J.; Gordon, Wendy R.

In: eLife, Vol. 8, e46983, 01.06.2019.

Research output: Contribution to journalArticle

@article{0d55409954854bceb05c5e14886412bb,
title = "A toolkit for studying cell surface shedding of diverse transmembrane receptors",
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.",
author = "Hayward, {Amanda N.} and Aird, {Eric J.} and Gordon, {Wendy R}",
year = "2019",
month = "6",
day = "1",
doi = "10.7554/eLife.46983",
language = "English (US)",
volume = "8",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",

}

TY - JOUR

T1 - A toolkit for studying cell surface shedding of diverse transmembrane receptors

AU - Hayward, Amanda N.

AU - Aird, Eric J.

AU - Gordon, Wendy R

PY - 2019/6/1

Y1 - 2019/6/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85068487960&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85068487960&partnerID=8YFLogxK

U2 - 10.7554/eLife.46983

DO - 10.7554/eLife.46983

M3 - Article

C2 - 31172946

AN - SCOPUS:85068487960

VL - 8

JO - eLife

JF - eLife

SN - 2050-084X

M1 - e46983

ER -