Mapping proteolytic neo-N termini at the surface of living cells.

Proc Natl Acad Sci U S A

Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143;

Published: February 2021

N terminomics is a powerful strategy for profiling proteolytic neo-N termini, but its application to cell surface proteolysis has been limited by the low relative abundance of plasma membrane proteins. Here we apply plasma membrane-targeted subtiligase variants (subtiligase-TM) to efficiently and specifically capture cell surface N termini in live cells. Using this approach, we sequenced 807 cell surface N termini and quantified changes in their abundance in response to stimuli that induce proteolytic remodeling of the cell surface proteome. To facilitate exploration of our datasets, we developed a web-accessible Atlas of Subtiligase-Captured Extracellular N Termini (ASCENT; http://wellslab.org/ascent). This technology will facilitate greater understanding of extracellular protease biology and reveal neo-N termini biomarkers and targets in disease.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7923363PMC
http://dx.doi.org/10.1073/pnas.2018809118DOI Listing

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