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Differently Tagged Probes for Protein Profiling of Mitochondria
Author(s) -
Dong Jia,
Hong Danqi,
Lang Wenjie,
Huang Jintao,
Qian Linghui,
Zhu Qing,
Li Lin,
Ge Jingyan
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800735
Subject(s) - mitochondrion , organelle , proteome , proteomics , protein subcellular localization prediction , mitochondrial matrix , microbiology and biotechnology , cysteine , biology , click chemistry , inner mitochondrial membrane , cell fractionation , chemistry , biochemistry , cytosol , biophysics , membrane , gene , combinatorial chemistry , enzyme
The mitochondrion is one of the most important organelles in the eukaryotic cell. Characterization of the mitochondrial proteome is a prerequisite for understanding its cellular functions at the molecular level. Here we report a proteomics method based on mitochondrion‐targeting groups and click chemistry. In our strategy, three different mitochondrion‐targeting moieties were each augmented with a clickable handle and a cysteine‐reactive group. Fluorescence‐based bioimaging and fractionation experiments clearly showed that most signals arising from the labels were localized in the mitochondria of cells, as a result of covalent attachment between probe and target proteins. The three probes had distinct profiling characteristics. Furthermore, we successfully identified more than two hundred mitochondrial proteins. The results showed that different mitochondrion‐targeting groups targeted distinct proteins with partial overlap. Most of the labeled proteins were localized in the mitochondrial matrix and inner mitochondrial membrane. Our results provide a tool for chemoproteomic analysis of mitochondrion‐related proteins.