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

ABSTRACT: 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.

SUBMITTER: Weeks AM

PROVIDER: S-EPMC7923363 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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

Weeks Amy M AM Byrnes James R JR Lui Irene I Wells James A JA

Proceedings of the National Academy of Sciences of the United States of America 20210201 8

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 prote ...[more]

PMID: 33536314

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