Journal
ANALYTICAL CHEMISTRY
Volume 90, Issue 15, Pages 9576-9582Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.8b02426
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Funding
- Ministry of Science and Technology of China [2016YFA0501500]
- National Science Foundation of China [21778004, 21521003, 81490741]
- 1000 Talents Plan Young Investigator Award
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Activity-based protein profiling (ABPP) has emerged as a powerful functional chemoproteomic strategy which enables global profiling of proteome reactivity toward bioactive small molecules in complex biological and/or pathological processes. To quantify the degree of reactivity in a site-specific manner, an isotopic tandem orthogonal proteolysis (isoTOP)-ABPP strategy has been developed; however, the high cost and long workflow associated with the synthesis of isotopically labeled cleavable tags limit its wide use. Herein, we combined reductive dimethyl labeling with TOP-ABPP to develop a fast, affordable, and efficient method, termed rdTOP-ABPP, for quantitative chemical proteomics with site-specific precision and triplex quantification. The rdTOP-ABPP method shows high accuracy and precision, good reproducibility, and better capacity for site identification and quantification and is highly compatible with many commercially available cleavable tags. We demonstrated the power of rdTOP-ABPP by profiling the target of (1S,3R)-RSL3, a canonical inducer for cell ferroptosis, and provided the first global portrait of its proteome reactivity in a quantitative and site-specific manner.
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