A Quantitative Chemical Proteomics Approach to Profile the Specific Cellular Targets of Andrographolide, a Promising Anticancer Agent That Suppresses Tumor Metastasis

作者：Wang Jigang; Tan Xing Fei; Van Sang Nguyen; Yang Peng; Zhou Jing; Gao Mingming; Li Zhengjun; Lim Teck Kwang; He Yingke; Ong Chye Sun; Lay Yifei; Zhang Jianbin; Zhu Guili; Lai Siew Li; Ghosh Dipanjana; Mok Yu Keung; Shen Han Ming; Lin Qingsong^*

来源：MOLECULAR & CELLULAR PROTEOMICS, 2014, 13(3): 876-886.

DOI：10.1074/mcp.M113.029793

Drug target identification is a critical step toward understanding the mechanism of action of a drug, which can help one improve the drug's current therapeutic regime and expand the drug's therapeutic potential. However, current in vitro affinity-chromatography-based and in vivo activity-based protein profiling approaches generally face difficulties in discriminating specific drug targets from nonspecific ones. Here we describe a novel approach combining isobaric tags for relative and absolute quantitation with clickable activity-based protein profiling to specifically and comprehensively identify the protein targets of andrographolide (Andro), a natural product with known anti-inflammation and anti-cancer effects, in live cancer cells. We identified a spectrum of specific targets of Andro, which furthered our understanding of the mechanism of action of the drug. Our findings, validated through cell migration and invasion assays, showed that Andro has a potential novel application as a tumor metastasis inhibitor. Moreover, we have unveiled the target binding mechanism of Andro with a combination of drug analog synthesis, protein engineering, and mass-spectrometry-based approaches and determined the drug-binding sites of two protein targets, NF-B and actin.

• 出版日期2014-3