Spitz Jean-Alexis, Polard Valérie, Maksimenko Andréi, Subra Frédéric, Baratti-Elbaz Catherine, Méallet-Renault Rachel, Pansu Robert B, Tauc Patrick, Auclair Christian
LBPA, Institut d'Alembert, ENS Cachan, CNRS, UniverSud, 61 av President Wilson, F-94230 Cachan, France.
Anal Biochem. 2007 Aug 1;367(1):95-103. doi: 10.1016/j.ab.2007.04.001. Epub 2007 Apr 5.
To study cellular actin dynamics, a cell-free assay based on fluorescence anisotropy was developed. Using G-actin-Alexa as a probe, we found that anisotropy enhancement reflects F-actin elongation. Anisotropy enhancement varies with the concentration of magnesium and calcium cations and with ethylenediaminetetraacetate or well-known effectors of the polymerization. This assay gives the overall status of actin dynamics in cell extracts which are the closest conditions to in vivo, implying most of the regulating proteins that are missing in purified actin measurements. It can be used in a large-scale screening for chemical compounds which modulate actin polymerization.
为了研究细胞肌动蛋白动力学，开发了一种基于荧光各向异性的无细胞检测方法。使用G-肌动蛋白- Alexa作为探针，我们发现各向异性增强反映了F-肌动蛋白的伸长。各向异性增强随镁离子和钙离子的浓度以及乙二胺四乙酸或众所周知的聚合效应剂而变化。该检测方法给出了细胞提取物中肌动蛋白动力学的整体状态，这是最接近体内的条件，意味着在纯化肌动蛋白测量中缺失的大多数调节蛋白。它可用于大规模筛选调节肌动蛋白聚合的化合物。
