Cell-Morphology Profiling of a Natural Product Library Identifies Bisebromoamide and Miuraenamide A as Actin Filament Stabilizers
Cell-Morphology Profiling of a Natural Product Library Identifies Bisebromoamide and Miuraenamide A as Actin Filament Stabilizers
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DOI：
10.1021/cb1003459
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发表时间：
2011-05-01
影响因子：
4
通讯作者：
Uesugi, Motonari
中科院分区：
文献类型：
--
作者：
Sumiya, Eriko;Shimogawa, Hiroki;Uesugi, Motonari
天然产物提供了丰富的生物工具来源，但阐明其分子靶标仍然具有挑战性。在这里，我们报告的细胞形态分析的天然产物库，允许识别bisebromoamide和miuraenamide A作为肌动蛋白丝稳定剂。自动化高内容图像分析表明，这两种结构不同的海洋天然产物诱导HeLa细胞的形态变化类似于已知的肌动蛋白稳定化合物诱导的那些。Bisebromoamide和miuraenamide A在体外稳定了肌动蛋白丝，荧光素结合的bisebromoamide特异性地定位于细胞中的肌动蛋白丝。细胞形态学分析也被用于鉴定从海绵提取物中的肌动蛋白稳定或不稳定的天然产物，从而分离出pectenotoxin-2和lyngbyabellin C。总体而言，结果表明，核和细胞形状的高内容成像提供了一个敏感的方便的方法，用于检测和分离分子，目标肌动蛋白。
Natural products provide a rich source of biological tools, but elucidating their molecular targets remains challenging. Here we report a cell morphological profiling of a natural product library, which permitted the identification of bisebromoamide and miuraenamide A as actin filament stabilizers. Automated high-content image analysis showed that these two structurally distinct marine natural products induce morphological changes in HeLa cells similar to those induced by known actin-stabilizing compounds. Bisebromoamide and miuraenamide A stabilized actin filaments in vitro, and fluorescein-conjugated bisebromoamide localized specifically to actin filaments in cells. Cell morphological profiling was also used to identify actin-stabilizing or -destabilizing natural products from marine sponge extracts, leading to the isolation of pectenotoxin-2 and lyngbyabellin C. Overall, the results demonstrate that high-content imaging of nuclei and cell shapes offers a sensitive convenient method for detecting and isolating molecules that target actin.