Degradation-Based Protein Profiling: A Case Study of Celastrol.

Area of Science:

• Drug Discovery
• Chemical Biology
• Proteomics

Background:

• Natural products are vital for drug discovery but often have unclear targets and toxicity.
• Celastrol, a traditional Chinese medicine component, has diverse activities but an unknown mechanism.
• Existing target identification methods face limitations.

Purpose of the Study:

• To develop and validate a novel Degradation-based Protein Profiling (DBPP) strategy for identifying natural product targets.
• To elucidate the molecular targets of celastrol using the DBPP approach.
• To demonstrate the efficiency and cost-effectiveness of DBPP for multi-target identification.

Main Methods:

• Development of a Degradation-based Protein Profiling (DBPP) strategy.
• Integration of PROteolysis TArgeting Chimeras (PROTAC) technology with quantitative proteomics and Immunoprecipitation-Mass Spectrometry (IP-MS).
• Utilizing a toolbox of degraders for target identification, including the development of the first glycosidase degrader.

Main Results:

• Successful identification of known celastrol targets: IKKβ, PI3Kα, and CIP2A.
• Discovery of potential new celastrol targets: CHK1, OGA, and ERCC6L.
• Achieved efficient and cost-effective multi-target identification of celastrol using a mixed PROTAC toolbox.

Conclusions:

• The DBPP strategy offers a powerful new approach for natural product target identification.
• This method complements existing strategies, accelerating pharmaceutical research and development.
• DBPP facilitates the discovery of both known and novel targets, enhancing drug development pipelines.