Fluorescent ligand fishing combination with in-situ imaging and characterizing to screen Hsp 90 inhibitors from Curcuma longa L. based on InP/ZnS quantum dots embedded mesoporous nanoparticles
Fluorescent ligand fishing combination with in-situ imaging and characterizing to screen Hsp 90 inhibitors from Curcuma longa L. based on InP/ZnS quantum dots embedded mesoporous nanoparticles
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基于 InP/ZnS 量子点嵌入介孔纳米颗粒的荧光配体捕获与原位成像和表征相结合筛选姜黄中的 Hsp 90 抑制剂
DOI：
10.1016/j.talanta.2017.09.035
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发表时间：
2018-02-01
期刊：
影响因子：
6.1
通讯作者：
Li, Wei
中科院分区：
文献类型：
--
作者：
Hu, Yue;Fu, Anchen;Li, Wei
虽然配体钓鱼已被证明是一种有效的技术，用于识别生物活性成分的复杂混合物，如天然产物，它不能应用于生物医学图像处理。本文提出了一种特异性荧光配体钓鱼结合原位成像的方法，用于从复杂基质姜黄中鉴定热休克蛋白90（Hsp 90）抑制剂，使用N-末端固定化的Hsp 90 α官能化的InP/ZnS量子凝块嵌入的介孔纳米颗粒（即Hsp 90 α（NT）-HspDNs）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）对捕获的配体进行鉴定。此外，原位成像共聚焦激光扫描显微镜（CLSM）应用于评估鱼配体的生物活性诱导的HeLa细胞凋亡形态学的影响。MTT法验证了配体的生物活性，分子对接结果进一步验证了配体与蛋白质的结合方式。从姜黄中筛选并鉴定了12个Hsp 90潜在抑制剂配体.粗提物该方法具有上级的特异性筛选效率和实时可视化原位成像的优点，为开发其他植物来源的Hsp 90抑制剂提供了一个快速、可靠的平台，为天然产物中生物活性分子的发现提供了一个新的途径。
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 90 alpha functionalized InP/ZnS quantum clots embedded mesoporous nanoparticles (i.e. Hsp 90 alpha (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 Tonga L. crude extracts. The proposed approach based on Hsp 90a 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.