Assessment of cellular actin dynamics by measurement of fluorescence anisotropy

Jean-Alexis Spitz (1, 2) , Valérie Polard (2, 3) , Andrei V. Maksimenko (2, 3) , Frédéric Subra (2, 3) , Catherine Baratti-Elbaz (2, 3) , Rachel Méallet-Renault (1, 2) , Robert Bernard Pansu (1, 2) , Patrick Tauc (2, 3) , Christian Auclair (2, 3)

1 PPSM - Laboratoire de Photophysique et Photochimie Supramoléculaires et Macromoléculaires
2 IDA - Institut d'Alembert
3 LBPA - Laboratoire de Biologie et de Pharmacologie Appliquée

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.

Mots clés: Actin cytoskeleton; Fluorescence anisotropy; Polymerization

Domaines: Chimie-Physique [physics.chem-ph]

Jean-Alexis Spitz, Valérie Polard, Andrei V. Maksimenko, Frédéric Subra, Catherine Baratti-Elbaz, et al.. Assessment of cellular actin dynamics by measurement of fluorescence anisotropy. Analytical Biochemistry, 2007, 367, pp.95-103. ⟨10.1016/j.ab.2007.04.001⟩. ⟨hal-00162079⟩