Versatile fluorescent probes for actin filaments based on the actin-binding domain of Utrophin
Versatile fluorescent probes for actin filaments based on the actin-binding domain of Utrophin
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DOI：
10.1002/cm.20226
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发表时间：
2007-11-01
影响因子：
--
通讯作者：
Bement, William M.
中科院分区：
文献类型：
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作者：
Burkel, Brian M.;von Dassow, George;Bement, William M.
肌动蛋白丝（F-actin）是蛋白质聚合物，其经历快速组装和拆卸并控制从力产生到信号转导调节的大量细胞过程。因此，F-肌动蛋白的成像已经成为生物学家寻求了解细胞和组织功能的一个越来越重要的目标。然而，大多数用于活细胞中的F-肌动蛋白成像的可用手段遭受一个或多个生物学或实验缺点。在这里，我们描述了荧光F-肌动蛋白探针的基础上的肌营养蛋白（Utr-CH），结合F-肌动蛋白，而不稳定它在体外的钙调蛋白同源域。我们发现，这些探针忠实地报告的F-肌动蛋白在活细胞和固定的细胞中的分布，区分稳定和动态的F-肌动蛋白，并没有明显的影响，关键取决于肌动蛋白组装和拆卸的平衡过程。
Actin filaments (F-actin) are protein polymers that undergo rapid assembly and disassembly and control an enormous variety of cellular processes ranging from force production to regulation of signal transduction. Consequently, imaging of F-actin has become an increasingly important goal for biologists seeking to understand how cells and tissues function. However, most of the available means for imaging F-actin in living cells suffer from one or more biological or experimental shortcomings. Here we describe fluorescent F-actin probes based on the calponin homology domain of utrophin (Utr-CH), which binds F-actin without stabilizing it in vitro. We show that these probes faithfully report the distribution of F-actin in living and fixed cells, distinguish between stable and dynamic F-actin, and have no obvious effects on processes that depend critically on the balance of actin assembly and disassembly.