Directed evolution of APEX2 for electron microscopy and proximity labeling
Name
Ting_Directed evolution.pdf
Size
2.08 MB
Format
Adobe PDF
Checksum (MD5)
8b372639cd48c0af1a4db0f034db291d
Author(s)
Kamer, Kimberli J
Deerinck, Thomas J
Ellisman, Mark H
Mootha, Vamsi K
Lam, Stephanie Shih-Min
Martell, Jeffrey Daniel
Ting, Alice Y
Date Issued
November 2014
Journal
Nature Methods
Publisher
Nature Publishing Group
Citation
Lam, Stephanie S; Martell, Jeffrey D; Kamer, Kimberli J et al. "Directed evolution of APEX2 for electron microscopy and proximity labeling." Nature Methods 12, 1: 51–54 (January 2015) © 2015 Nature America, Inc
Version
Author's final manuscript
Abstract
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.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/nmeth.3179
