Although ligand fishing has been shown to be an efficient technique for the identification of bioactive components from complex mixtures such as natural products, it cannot be applied to biomedical image processing. Herein, a specific fluorescent ligand fishing combined with in situ imaging approach is presented for the identification of heat shock protein 90 (Hsp 90) inhibitors from complex matrixes, Curcuma longa L., using N-terminus immobilized Hsp 90α functionalized InP/ZnS quantum dots embedded mesoporous nanoparticles (i.e. Hsp 90α (NT)-FQDNs) as extraction sorbents and fluorescent tracer. The fished ligands were identified by liquid chromatography time-of-flight/mass spectrometry (LC-TOF/MS) and gas chromatography-mass spectrometry (GC-MS). Moreover, in situ imaging by confocal laser scanning microscopy (CLSM) was applied for evaluating the effect of fished-ligands on bioactivity-induced apoptosis morphologically in HeLa cells. MTT assay verified the bioactivity of the ligands and molecular docking results further provided convincing information to verify the feasible binding mode between ligands and protein. Twelve ligands as potential Hsp 90 inhibitors were ultimately fished and identified from Curcuma longa L. crude extracts. The proposed approach based on Hsp 90α functionalized nanocomposites is superior in the combination of highly specific screening efficiency and concurrent visual in situ imaging, which could have great promise for the development of other plant-derived Hsp 90 inhibitors, and providing a rapid and reliable platform for discovering biologically active molecules in natural products.

尽管配体捕捞已被证明是从复杂混合物（例如天然产物）中鉴定生物活性成分的有效技术，但它不能应用于生物医学图像处理。本文中，提出了一种特定的荧光配体捕捞与原位成像方法相结合的方法，用于使用N末端固定化的Hsp90α功能化InP / ZnS量子点从复杂基质姜黄中鉴定热休克蛋白90（Hsp 90）抑制剂。嵌入中孔纳米颗粒（即Hsp90α（NT）-FQDNs）作为提取吸附剂和荧光示踪剂。通过液相色谱飞行时间/质谱（LC-TOF / MS）和气相色谱-质谱（GC-MS）鉴定所捕捞的配体。而且，共聚焦激光扫描显微镜（CLSM）的原位成像技术被用于评估鱼卵配体对HeLa细胞形态学上生物活性诱导的细胞凋亡的影响。MTT分析验证了配体的生物活性，分子对接结果进一步提供了令人信服的信息，以验证配体与蛋白质之间的可行结合方式。最终捕捞了十二种作为潜在Hsp 90抑制剂的配体，并从姜黄粗提取物中鉴定出了它们。基于Hsp90α功能化纳米复合材料的拟议方法在高度特异性筛选效率和同时进行原位视觉结合方面具有优势 成像技术，对于开发其他植物来源的Hsp 90抑制剂可能具有广阔的前景，并为发现天然产物中的生物活性分子提供了一个快速而可靠的平台。