Cell Cortex Composition and Homeostasis Resolved by Integrating Proteomics and Quantitative Imaging
Cell Cortex Composition and Homeostasis Resolved by Integrating Proteomics and Quantitative Imaging
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DOI：
10.1002/cm.21142
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发表时间：
2013-11-01
期刊：
影响因子：
2.9
通讯作者：
Paluch，Ewa K.
中科院分区：
文献类型：
--
作者：
Biro，Mate;Romeo，Yves;Paluch，Ewa K.
细胞肌动蛋白皮层是主要负责控制动物细胞形状的细胞骨架结构，因此在细胞分裂、迁移和组织形态发生中起着核心作用。由于缺乏可以独立于其他细胞器研究皮层的实验系统，对其组成、组装和稳态知之甚少。在这里，我们描述了新的工具来解决的组成和调节的皮层。我们报告并验证了一种基于细胞泡分离的皮层纯化方案。质谱分析纯化的皮质提供了皮质成分的第一个广泛的列表。为了评估鉴定蛋白的功能，我们设计了一种自动成像分析，用于精确定量皮质肌动球蛋白组装动力学。我们显示了在确定的皮层成分谱枯竭后，皮层组装动力学的微妙变化。我们广泛应用的集成方法为在系统水平上研究肌动球蛋白皮层及其在形态发生过程中的调节铺平了道路。©2013 Wiley期刊公司
The cellular actin cortex is the cytoskeletal structure primarily responsible for the control of animal cell shape and as such plays a central role in cell division, migration, and tissue morphogenesis. Due to the lack of experimental systems where the cortex can be investigated independently from other organelles, little is known about its composition, assembly, and homeostasis. Here, we describe novel tools to resolve the composition and regulation of the cortex. We report and validate a protocol for cortex purification based on the separation of cellular blebs. Mass spectrometry analysis of purified cortices provides a first extensive list of cortical components. To assess the function of identified proteins, we design an automated imaging assay for precise quantification of cortical actomyosin assembly dynamics. We show subtle changes in cortex assembly dynamics upon depletion of the identified cortical component profilin. Our widely applicable integrated method paves the way for systems‐level investigations of the actomyosin cortex and its regulation during morphogenesis. © 2013 Wiley Periodicals, Inc.