Assessment of cellular actin dynamics by measurement of fluorescence anisotropy

被引:6
作者
Spaz, Jean-Alexis
Polard, Valerie
Maksimenko, Andrei
Subra, Frederic
Baratti-Elbaz, Catherine
Meallet-Renault, Rachel
Pansu, Robert B.
Tauc, Patrick
Auclair, Christian
机构
[1] Univ Sud, CNRS, ENS Cachan, LBPA,Inst Alembert, F-94230 Cachan, France
[2] Univ Sud, CNRS, ENS Cachan, PPSM,Inst Alembert, F-94230 Cachan, France
[3] BioAlliance Pharma, F-75015 Paris, France
关键词
actin cytoskeleton; fluorescence anisotropy; polymerization;
D O I
10.1016/j.ab.2007.04.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2007 Elsevier Inc, All rights reserved.
引用
收藏
页码:95 / 103
页数:9
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