High-Content Imaging Platform to Discover Chemical Modulators of Plasma Membrane Rafts.
High-Content Imaging Platform to Discover Chemical Modulators of Plasma Membrane Rafts.
复制标题
发现质膜筏化学调节剂的高含量成像平台。
DOI：
10.1021/acscentsci.1c01058
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发表时间：
2022-03-23
影响因子：
18.2
通讯作者：
Kenworthy AK
中科院分区：
文献类型：
--
作者：
Fricke N;Raghunathan K;Tiwari A;Stefanski KM;Balakrishnan M;Waterson AG;Capone R;Huang H;Sanders CR;Bauer JA;Kenworthy AK
质膜组织深刻影响细胞功能。这种组织的一个众所周知的机制是通过膜筏中不同的脂质和蛋白质的纳米级聚集。尽管膜筏具有重要的生理意义，但它仍然是膜组织中一个难以研究的方面，部分原因是缺乏化学工具来实验调节其性质。目前也缺乏选择性靶向治疗目的的方法。为了解决这些问题，我们开发了一种高通量屏幕和附带的图像分析管道，以识别增强或抑制筏形成的小分子。以细胞源性巨质膜囊泡为实验平台。使用生物活性脂质库的原理验证筛选表明，该方法是稳健的，能够验证已建立的筏状调节剂，包括C6-和c8 -神经酰胺、米特氟辛和表没食子儿茶素没食子酸酯，以及识别新的。我们在这里描述的平台代表了一个强大的工具，发现新的化学方法来操纵筏及其组件。我们提出了一种基于显微镜的方法来识别细胞源性质膜中作为筏调节剂的小分子。我们的软件有利于膜囊的高含量分析。
Plasma membrane organization profoundly impacts cellular functionality. A well-known mechanism underlying this organization is through nanoscopic clustering of distinct lipids and proteins in membrane rafts. Despite their physiological importance, rafts remain a difficult-to-study aspect of membrane organization, in part because of the paucity of chemical tools to experimentally modulate their properties. Methods to selectively target rafts for therapeutic purposes are also currently lacking. To tackle these problems, we developed a high-throughput screen and an accompanying image analysis pipeline to identify small molecules that enhance or inhibit raft formation. Cell-derived giant plasma membrane vesicles were used as the experimental platform. A proof-of-principle screen using a bioactive lipid library demonstrates that this method is robust and capable of validating established raft modulators including C6- and C8-ceramide, miltefosine, and epigallocatechin gallate as well as identifying new ones. The platform we describe here represents a powerful tool to discover new chemical approaches to manipulate rafts and their components. We present a microscopy-based method to identify small molecules that function as raft modulators in cell-derived plasma membranes. Our software facilitates high-content analysis of membrane vesicles.
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影响因子：
4
作者：
Ingolfsson, Helgi I.;Thakur, Pratima;Herold, Karl F.;Hobart, E. Ashley;Ramsey, Nicole B.;Periole, Xavier;de Jong, Djurre H.;Zwama, Martijn;Yilmaz, Duygu;Hall, Katherine;Maretzky, Thorsten;Hemmings, Hugh C., Jr.;Blobel, Carl;Marrink, Siewert J.;Kocer, Armagan;Sack, Jon T.;Andersen, Olaf S.
通讯作者：
Andersen, Olaf S.
影响因子：
3
作者：
GHOSE, AK;CRIPPEN, GM
通讯作者：
CRIPPEN, GM
影响因子：
3.9
作者：
Neves, Ana Rute;Nunes, Claudia;Reis, Salette
通讯作者：
Reis, Salette
影响因子：
3.4
作者：
Cornell CE;McCarthy NLC;Levental KR;Levental I;Brooks NJ;Keller SL
通讯作者：
Keller SL
影响因子：
3.4
作者：
Levental, Kandice R.;Lorent, Joseph H.;Levental, Ilya
通讯作者：
Levental, Ilya