A PEROXO-Tag enables rapid isolation of peroxisomes from human cells
Author(s)
Ray, G Jordan; Boydston, Elizabeth A; Shortt, Emily; Wyant, Gregory A; Lourido, Sebastian; Chen, Walter W; Sabatini, David M; ... Show more Show less
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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.
Date issued
2020Department
Whitehead Institute for Biomedical Research; Howard Hughes Medical Institute; Koch Institute for Integrative Cancer Research at MIT; Massachusetts Institute of Technology. Department of BiologyJournal
iScience
Publisher
Elsevier BV