Fluorescence image screening for chemical compounds modifying cholesterol metabolism and distribution[S]
Fluorescence image screening for chemical compounds modifying cholesterol metabolism and distribution[S]
复制标题
改变胆固醇代谢和分布的化合物的荧光图像筛选[S]
DOI：
--
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发表时间：
2011
影响因子：
6.5
通讯作者：
Toshihide Kobayashi
中科院分区：
文献类型：
--
作者：
R. Ishitsuka;Tamio Saito;H. Osada;Y. Ohno‐Iwashita;Toshihide Kobayashi
一种使用无毒胆固醇结合蛋白θ毒素结构域4 （D4）的自动荧光显微镜分析方法被开发出来，以识别改变细胞内胆固醇代谢和分布的化合物。利用这种方法，我们筛选了1,056个化合物，并鉴定出35个化合物可以降低D4与细胞表面的结合。其中，已有8种化合物被报道可以改变胆固醇的生物合成或细胞内分布。对其余27个hit化合物进行生化和组织化学分析。用另一种荧光胆固醇探针filipin对细胞进行染色，发现17种化合物在晚期核内体中积累了胆固醇。5种化合物降低了胆固醇的生物合成，2种化合物抑制了D4与细胞膜的结合。这种基于胆固醇特异性探针D4与生化分析相结合的视觉筛选方法，是一种基于细胞的、敏感的识别新化合物和改变胆固醇分布和代谢的技术。此外，它还适用于药物发现的高通量分析。
An automated fluorescence microscopy assay using a nontoxic cholesterol binding protein, θ toxin domain 4, (D4), was developed in order to identify chemical compounds modifying intracellular cholesterol metabolism and distribution. Using this method, we screened a library of 1,056 compounds and identified 35 compounds that decreased D4 binding to the cell surface. Among them, 8 compounds were already reported to alter the biosynthesis or the intracellular distribution of cholesterol. The remaining 27 hit compounds were further analyzed biochemically and histochemically. Cell staining with another fluorescent cholesterol probe, filipin, revealed that 17 compounds accumulated cholesterol in the late endosomes. Five compounds decreased cholesterol biosynthesis, and two compounds inhibited the binding of D4 to the membrane. This visual screening method, based on the cholesterol-specific probe D4 in combination with biochemical analyses, is a cell-based, sensitive technique for identifying new chemical compounds and modifying cholesterol distribution and metabolism. Furthermore, it is suitable for high-throughput analysis for drug discovery.
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DOI：
10.1016/j.bbalip.2009.03.002
发表时间：
2009-07
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Mesmin B;Maxfield FR
通讯作者：
Maxfield FR
影响因子：
2.9
作者：
R. Sexton;S. Panini;F. Azran;H. Rudney
通讯作者：
R. Sexton;S. Panini;F. Azran;H. Rudney
影响因子：
7.5
作者：
Maxfield FR;van Meer G
通讯作者：
van Meer G
Niemann-Pick C 型和正常人成纤维细胞中溶酶体胆固醇的动态。
影响因子：
6.5
作者：
Lange,Yvonne;Ye,Jin;Rigney,Mike;Steck,TheodoreL
通讯作者：
Steck,TheodoreL