Direct determination of actin polarity in the cell.
Direct determination of actin polarity in the cell.
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DOI：
10.1016/j.jmb.2012.03.015
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发表时间：
2012-06-22
影响因子：
5.6
通讯作者：
Maeda, Yuichiro
中科院分区：
文献类型：
--
作者：
Narita, Akihiro;Mueller, Jan;Urban, Edit;Vinzenz, Marlene;Small, J. Victor;Maeda, Yuichiro
肌动蛋白丝是一种极性结构，具有快速生长的正端和缓慢生长的负端。根据它们在细胞内的排列，平行排列或反平行排列，它们可以分别以突起或收缩的形式发挥作用。因此，确定亚细胞区室中的肌动蛋白丝极性是确定其局部功能所必需的。肌球蛋白结合以前是测定极性的唯一方法。在这里，我们报道了在没有肌凝蛋白结合的情况下，细胞中肌动蛋白丝极性的第一次直接测定。采用电子断层扫描和丝状复合体的单粒子分析对板足细胞骨架进行了阴性染色。细胞骨架染色结果证实，所有面向细胞膜的肌动蛋白丝端均为倒刺端。一般来说，这种方法应该适用于肌动蛋白细胞骨架断层扫描中肌动蛋白极性的分析。在没有肌凝蛋白修饰的情况下，直接测定细胞中的肌动蛋白极性。►板足阴性染色的电子断层图。►对丝状复合物采用单粒子分析。
Actin filaments are polar structures that exhibit a fast growing plus end and a slow growing minus end. According to their organization in cells, in parallel or antiparallel arrays, they can serve, respectively, in protrusions or in contractions. The determination of actin filament polarity in subcellular compartments is therefore required to establish their local function. Myosin binding has previously been the sole method of polarity determination. Here, we report the first direct determination of actin filament polarity in the cell without myosin binding. Negatively stained cytoskeletons of lamellipodia were analyzed by adapting electron tomography and a single particle analysis for filamentous complexes. The results of the stained cytoskeletons confirmed that all actin filament ends facing the cell membrane were the barbed ends. In general, this approach should be applicable to the analysis of actin polarity in tomograms of the actin cytoskeleton. ► Actin polarity in the cell was directly determined without myosin decoration. ► Electron tomograms of negatively stained lamellipodia. ► A single particle analysis for filamentous complexes was employed.
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影响因子：
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作者：
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通讯作者：
Small, J. Victor
影响因子：
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