A PEROXO-Tag enables rapid isolation of peroxisomes from human cells
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1-s2.0-S2589004220302947-main.pdf
Description
Published version
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1.6 MB
Format
Adobe PDF
Checksum (MD5)
b6d5d338a06386e8a04357910db80e8d
Author(s)
Ray, G Jordan
Boydston, Elizabeth A
Shortt, Emily
Wyant, Gregory A
Lourido, Sebastian
Chen, Walter W
Sabatini, David M
Date Issued
2020
Journal
iScience
Publisher
Elsevier BV
Version
Final published version
Abstract
Peroxisomes are metabolic organelles that perform a diverse array of critical functions in human physiology. Traditional isolation methods for peroxisomes can take more than 1 h to complete and can be laborious to implement. To address this, we have now extended our prior work on rapid organellar isolation to peroxisomes via the development of a peroxisomally localized 3XHA epitope tag (“PEROXO-Tag”) and associated immunoprecipitation (“PEROXO-IP”) workflow. Our PEROXO-IP workflow has excellent reproducibility, is easy to implement, and achieves highly rapid (~10 min post homogenization) and specific isolation of human peroxisomes, which we characterize here via proteomic profiling. By offering speed, specificity, reproducibility, and ease of use, the PEROXO-IP workflow should facilitate studies on the biology of peroxisomes.
MIT Department
Whitehead Institute for Biomedical Research
Howard Hughes Medical Institute
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/J.ISCI.2020.101109
