RT Journal Article
SR Electronic
T1 SNAPS: A toolkit for studying cell surface shedding of diverse transmembrane receptors
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 436592
DO 10.1101/436592
A1 Amanda N. Hayward
A1 Eric J. Aird
A1 Wendy R. Gordon
YR 2018
UL http://biorxiv.org/content/early/2018/10/24/436592.abstract
AB Proteolysis of transmembrane receptors is a critical cellular communication mechanism dysregulated in many diseases, yet decoding proteolytic regulation mechanisms of the estimated 400 receptors shed from the cell surface has been hindered by difficulties in controlling stimuli and unknown fates of cleavage products. Notch proteolytic regulation is a notable exception, where decades of study have revealed that intercellular forces drive exposure of a cryptic protease site within a juxtamembrane “proteolytic switch” domain to activate transcriptional programs inside the cell. Thus, 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. Here, we identify several new proteolytic switches among receptors with structural homology to Notch. We demonstrate that SNAPS can detect shedding in chimeras of diverse cell surface receptors, leading to new, testable hypotheses. Finally, we establish that the assay can be used to measure modulation of proteolysis by potential therapeutics.