title: Assessment of cellular actin dynamics by measurement of fluorescence anisotropy.
abstract: 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.
authors.affiliation: LBPA, Institut d'Alembert, ENS Cachan, CNRS, UniverSud, 61 av President Wilson, F-94230 Cachan, France.
authors.forename: Jean-Alexis
authors.initials: JA
authors.lastname: Spitz
authors.forename: Valérie
authors.initials: V
authors.lastname: Polard
authors.forename: Andréi
authors.initials: A
authors.lastname: Maksimenko
authors.forename: Frédéric
authors.initials: F
authors.lastname: Subra
authors.forename: Catherine
authors.initials: C
authors.lastname: Baratti-Elbaz
authors.forename: Rachel
authors.initials: R
authors.lastname: Méallet-Renault
authors.forename: Robert B
authors.initials: RB
authors.lastname: Pansu
authors.forename: Patrick
authors.initials: P
authors.lastname: Tauc
authors.forename: Christian
authors.initials: C
authors.lastname: Auclair
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D002413:Cations, Divalent / Q000378:metabolism; D002471:Cell Transformation, Neoplastic; D002474:Cell-Free System; D002614:Chelating Agents; D003600:Cytosol / Q000378:metabolism; D047630:Depsipeptides / Q000494:pharmacology; D005454:Fluorescence Polarization / Q000379:methods*; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D051379:Mice; D041681:NIH 3T3 Cells; D011817:Rabbits; D013388:Succinimides; D013816:Thermodynamics
chemical_list: D000199:Actins; C492694:Alexa Fluor 488 carboxylic acid succinimidyl ester; D002413:Cations, Divalent; D002614:Chelating Agents; D047630:Depsipeptides; D005456:Fluorescent Dyes; D013388:Succinimides; C422265:dolastatin 11; C057531:jasplakinolide