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An in situ ascorbate peroxidase 2-based labeling method and its application in detecting exosome proteins released from mouse proximal tubule cells under oxidative stress

ABSTRACT: Here, we developed an in situ labeling approach to identify exosome-loaded proteins, which bypasses the traditional exosome isolation steps. In this method, an engineered ascorbate peroxidase targeting to endosomes-destined exosomes was used to label proteins loaded inside exosomes released from kidney proximal tubule-derived cells. Mass spectrometry analysis of the kidney proximal tubule-derived cells identified oxidative stress-induced exosome proteome alteration.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Epithelial Cell, Kidney

DISEASE(S): Disease Free

SUBMITTER: Sang-Ho Kwon

LAB HEAD: Sang-Ho Kwon

PROVIDER: PXD025852 | Pride | 2022-08-12

REPOSITORIES: Pride

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Ascorbate peroxidase-mediated in situ labelling of proteins in secreted exosomes.

Lee Byung Rho BR Lee Tae Jin TJ Oh Sekyung S Li Chenglong C Song Jin-Hyuk A JA Marshall Brendan B Zhi Wenbo W Kwon Sang-Ho SH

Journal of extracellular vesicles 20220601 6

The extracellular vesicle exosome mediates intercellular communication by transporting macromolecules such as proteins and ribonucleic acids (RNAs). Determining cargo contents with high accuracy will help decipher the biological processes that exosomes mediate in various contexts. Existing methods for probing exosome cargo molecules rely on a prior exosome isolation procedure. Here we report an in situ labelling approach for exosome cargo identification, which bypasses the exosome isolation step ...[more]

PMID: 35716063

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