A PEROXO-Tag Enables Rapid Isolation of Peroxisomes From Human Cells

iScience. 2020 May 22;23(5):101109. doi: 10.1016/j.isci.2020.101109. Epub 2020 Apr 28.

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.

Keywords: Biochemistry Methods; Biological Sciences; Cell Biology; Methodology in Biological Sciences.