Directed evolution of APEX2 for electron microscopy and proximity labeling

Directed evolution of APEX2 for electron microscopy and proximity labeling

APEX is an engineered peroxidase that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics. Because limited sensitivity precludes applications requiring low APEX expression, we used yeast-display evolution to improve its catalytic efficiency. APEX2 is fa...

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Bibliographic Details
Main Authors: Kamer, Kimberli J, Deerinck, Thomas J, Ellisman, Mark H, Mootha, Vamsi K, Lam, Stephanie Shih-Min, Martell, Jeffrey Daniel, Ting, Alice Y
Other Authors: Massachusetts Institute of Technology. Department of Chemistry
Format: Article
Language:en_US
Published: Nature Publishing Group 2017
Online Access:http://hdl.handle.net/1721.1/110613
https://orcid.org/0000-0002-2687-3470
https://orcid.org/0000-0002-8277-5226
Description
Summary:APEX is an engineered peroxidase that functions as an electron microscopy tag and a promiscuous labeling enzyme for live-cell proteomics. Because limited sensitivity precludes applications requiring low APEX expression, we used yeast-display evolution to improve its catalytic efficiency. APEX2 is far more active in cells, enabling the use of electron microscopy to resolve the submitochondrial localization of calcium uptake regulatory protein MICU1. APEX2 also permits superior enrichment of endogenous mitochondrial and endoplasmic reticulum membrane proteins.

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