Le Clainche Christophe, Carlier Marie-France
Laboratoire d'Enzymologie et Biochimie Structurales (LEBS), Centre National de la Recherche Scientifique (CNRS), Gif-sur-Yvette, France.
Curr Protoc Cell Biol. 2004 Oct;Chapter 12:12.7.1-12.7.20. doi: 10.1002/0471143030.cb1207s24.
Actin-based movement can be reconstituted by using microspheres functionalized with the enzymes N-WASP or ActA, which use the Arp2/3 complex and actin to catalyze the formation of a branched actin filament network that is maintained in rapid turnover by three proteins (capping protein, profilin, and ADF). The particles continuously initiate filament assembly at their surface and are propelled, mimicking bacteria or the leading edge of motile cells. This biomimetic assay offers advantages over approaches based on living cells and cell extracts, because the physical-chemical parameters are under control. The biomimetic motility assay offers the opportunity to test the function of proteins involved in signaling pathways or actin dynamics. It is a powerful tool to understand the physical mechanism of force production and has the potential to support high-throughput screens for drugs, inhibitors of motility, or therapeutic agents in metastatic states in which motility is impaired.
基于肌动蛋白的运动可以通过使用用N-WASP或ActA酶功能化的微球来重建，这些酶利用Arp2/3复合物和肌动蛋白催化分支肌动蛋白丝网络的形成，该网络由三种蛋白质（帽蛋白、肌动蛋白单体结合蛋白和肌动蛋白解聚因子）维持快速周转。这些颗粒在其表面持续启动丝状体组装并被推动，模拟细菌或运动细胞的前沿。这种仿生检测方法比基于活细胞和细胞提取物的方法具有优势，因为物理化学参数是可控的。仿生运动检测提供了测试参与信号通路或肌动蛋白动力学的蛋白质功能的机会。它是理解力产生物理机制的有力工具，并且有可能支持针对药物、运动抑制剂或转移性状态下运动受损的治疗剂的高通量筛选。
