This study introduces a novel "Degradation-based protein profiling (DBPP)" strategy, which combines PROteolysis Targeting Chimeras (PROTAC) technology with quantitative proteomics and Immunoprecipitation-Mass Spectrometry (IP-MS) techniques to identify multiple targets of natural products. Using celastrol as a case study, the researchers successfully identified known targets such as inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PI3Kα), as well as potential new targets like checkpoint kinase 1 (CHK1), O-GlcNAcase (OGA), and DNA excision repair protein ERCC-6-like (ERCC6L). The study also developed the first glycosidase degrader and demonstrated that a mixed PROTAC toolbox in quantitative proteomics significantly improved the efficiency and cost-effectiveness of target identification. The DBPP strategy is expected to complement existing target identification methods, accelerating drug discovery and advancing the pharmaceutical field.This study introduces a novel "Degradation-based protein profiling (DBPP)" strategy, which combines PROteolysis Targeting Chimeras (PROTAC) technology with quantitative proteomics and Immunoprecipitation-Mass Spectrometry (IP-MS) techniques to identify multiple targets of natural products. Using celastrol as a case study, the researchers successfully identified known targets such as inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PI3Kα), as well as potential new targets like checkpoint kinase 1 (CHK1), O-GlcNAcase (OGA), and DNA excision repair protein ERCC-6-like (ERCC6L). The study also developed the first glycosidase degrader and demonstrated that a mixed PROTAC toolbox in quantitative proteomics significantly improved the efficiency and cost-effectiveness of target identification. The DBPP strategy is expected to complement existing target identification methods, accelerating drug discovery and advancing the pharmaceutical field.