APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest
Name
Lee-2016-APEX Fingerprinting.pdf
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3.48 MB
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Checksum (MD5)
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Author(s)
Lee, Song-Yi
Kang, Myeong-Gyun
Park, Jong-Seok
Lee, Geunsik
Rhee, Hyun-Woo
Ting, Alice Y
Date Issued
May 2016
Journal
Cell Reports
Publisher
Elsevier
Citation
Lee, Song-Yi; Kang, Myeong-Gyun; Park, Jong-Seok; Lee, Geunsik; Ting, Alice Y. and Rhee, Hyun-Woo. “APEX Fingerprinting Reveals the Subcellular Localization of Proteins of Interest.” Cell Reports 15, 8 (May 2016): 1837–1847 © 2016 The Author(s)
Version
Final published version
Abstract
Deciphering the sub-compartmental location of a given protein of interest may help explain its physiological function, but it can be challenging to do using optical or biochemical methods. Imaging with electron microscopy (EM) can provide highly resolved mapping of proteins; however, EM requires complex sample preparation and a specialized facility. Here, we use engineered ascorbate peroxidase (APEX)-generated molecular labeling patterns to provide information regarding intracellular microenvironments in living cells. Using APEX labeling of specific proteins, we uncovered subcellular localization at sub-compartmental resolution and successfully elucidated the membrane protein topology of HMOX1 and sub-mitochondrial localization of recently identified mitochondrial proteins. This method can be expanded to confirm sub-mitochondrial localization and membrane topologies of previously identified mitochondrial proteins.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Creative Commons Attribution 4.0 International License
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DOI of Published Version
https://doi.org/10.1016/j.celrep.2016.04.064
