Xiangya Hospital, Central South University, Changsha 410008, China; College of Pharmacy, Guilin Medical University, Guilin 541004, China.
Research Center for Science, Guilin Medical University, Guilin 541004, China.
J Proteomics. 2018 Jun 30;182:1-11. doi: 10.1016/j.jprot.2018.04.025. Epub 2018 Apr 22.
Identification of the specific protein target(s) of a drug is a critical step in unraveling its mechanisms of action (MOA) in many natural products. Curcumol, isolated from well known Chinese medicinal plant Curcuma zedoary, has been shown to possess multiple biological activities. It can inhibit nasopharyngeal carcinoma (NPC) proliferation and induce apoptosis, but its target protein(s) in NPC cells remains unclear. In this study, we employed a mass spectrometry-based chemical proteomics approach reveal the possible protein targets of curcumol in NPC cells. Cellular thermal shift assay (CETSA), molecular docking and cell-based assay was used to validate the binding interactions. Chemical proteomics capturing uncovered that NCL is a target of curcumol in NPC cells, Molecular docking showed that curcumol bound to NCL with an -7.8 kcal/mol binding free energy. Cell function analysis found that curcumol's treatment leads to a degradation of NCL in NPC cells, and it showed slight effects on NP69 cells. In conclusion, our results providing evidences that NCL is a target protein of curcumol. We revealed that the anti-cancer effects of curcumol in NPC cells are mediated, at least in part, by NCL inhibition.
Many natural products showed high bioactivity, while their mechanisms of action (MOA) are very poor or completely missed. Understanding the MOA of natural drugs can thoroughly exploit their therapeutic potential and minimize their adverse side effects. Identification of the specific protein target(s) of a drug is a critical step in unraveling its MOA. Compound-centric chemical proteomics is a classic chemical proteomics approach which integrates chemical synthesis with cell biology and mass spectrometry (MS) to identify protein targets of natural products determine the drug mechanism of action, describe its toxicity, and figure out the possible cause of off-target. It is an affinity-based chemical proteomics method to identify small molecule-protein interactions through affinity chromatography approach coupled with mass spectrometry, has been conventionally used to identify target proteins and has yielded good results. Curcumol, has shown effective inhibition on Nasopharyngeal Carcinoma (NPC) Cells, interacted with NCL and then initiated the anti-tumor biological effect. This research demonstrated the effectiveness of chemical proteomics approaches in natural drugs molecular target identification, revealing and understanding of the novel mechanism of actions of curcumol is crucial for cancer prevention and treatment in nasopharynx cancer.
鉴定药物的特定蛋白靶标是揭示其在许多天然产物中的作用机制（MOA）的关键步骤。从著名的药用植物莪术中分离出的莪术醇已被证明具有多种生物活性。它可以抑制鼻咽癌（NPC）的增殖并诱导细胞凋亡，但 NPC 细胞中其靶标蛋白尚不清楚。在这项研究中，我们采用基于质谱的化学蛋白质组学方法来揭示莪术醇在 NPC 细胞中的可能靶标蛋白。细胞热转移分析（CETSA）、分子对接和基于细胞的测定用于验证结合相互作用。化学蛋白质组学捕获结果表明，NCL 是 NPC 细胞中莪术醇的靶标，分子对接表明莪术醇与 NCL 结合的结合自由能为-7.8 kcal/mol。细胞功能分析发现，莪术醇处理导致 NPC 细胞中 NCL 的降解，而对 NP69 细胞的影响较小。总之，我们的结果提供了证据表明 NCL 是莪术醇的靶标蛋白。我们揭示了莪术醇在 NPC 细胞中的抗癌作用至少部分是通过 NCL 抑制介导的。
许多天然产物表现出很高的生物活性，但其作用机制（MOA）很差或完全缺失。了解天然药物的 MOA 可以充分挖掘其治疗潜力，并最大程度地减少其不良反应。鉴定药物的特定蛋白靶标是揭示其 MOA 的关键步骤。基于化合物的化学蛋白质组学是一种经典的化学蛋白质组学方法，它将化学合成与细胞生物学和质谱（MS）相结合，以鉴定天然产物的靶标蛋白，确定药物的作用机制，描述其毒性，并找出可能的脱靶原因。它是一种基于亲和力的化学蛋白质组学方法，通过亲和层析方法与质谱相结合来鉴定小分子-蛋白质相互作用，已被常规用于鉴定靶标蛋白，并取得了良好的效果。莪术醇对鼻咽癌（NPC）细胞有明显的抑制作用，与 NCL 相互作用后启动抗肿瘤生物效应。本研究证明了化学蛋白质组学方法在天然药物分子靶标鉴定中的有效性，揭示和理解莪术醇的新作用机制对鼻咽癌的防治具有重要意义。
