Molecular analysis of microscopic ezrin dynamics by two-photon FRAP.
Ezrin plays a key role in coupling signal transduction to cortical cell organization. This actin-membrane linker undergoes a series of conformational changes that modulate its interactions with various partners and its localization in membrane or cytosolic pools. Its mobility and exchange rates within and between these two pools were assessed by two-photon fluorescence recovery after photobleaching in epithelial cell microvilli. Analysis of ezrin mutants with an altered actin-binding site revealed three ezrin membrane states of different mobilities and exchange properties, reflecting sequential association with membrane components and F-actin in the context of a fast overall turnover.
Coscoy
Sylvie
S
Laboratoires de Physico-Chimie,Unité Mixte de Recherche 168 and Morphogenèse et Signalisation Cellulaires, Unité Mixte de Recherche 144, Centre National de la Recherche Scientifique/Institut Curie, 26, Rue d'Ulm, 75248 Paris Cedex 05, France.
Waharte
François
F
Gautreau
Alexis
A
Martin
Marianne
M
Louvard
Daniel
D
Mangeat
Paul
P
Arpin
Monique
M
Amblard
Françis
F
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008027:Light; D008164:Luminescent Proteins / Q000378:metabolism; D008954:Models, Biological; D010750:Phosphoproteins / Q000737:chemistry* / Q000502:physiology*; D017785:Photons; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013552:Swine; D014162:Transfection; D000099149:Ezrin
D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D000099149:Ezrin