Cortical plaques Lambotin et al, J Cell Science 2005 Ezrin Actine (nid dabeille) Lambotin et al, J Cell Science 2005 Doulet et al, J Cell Biol 2006 Nécessaire à ladhésion "forte" en condition de flux (G. Mikaty, G. Dumenil & coll) PI3K GTPases
β-arrestins Lan Ma and Gang Pei, Journal of Cell Science 120, (2007) MapK Src Akt A BC
β-arrestins - SiRNA βarrestin 1/2 Actin Control siRNA βarrestin 1/2 siRNA 2,5 fold Expression is reduced by Beta arrestine 2 GFP ezrin Ezrin recruitment 0% 20% 40% 60% 80% 100% Control siRNA Arrestin 1+2 sirRNA FLNA sirRNAGRK2 sirRNA Ezrin
β-arrestins - SiRNA βarrestin 1/2 Filamin A GRK2 Actin Control siRNA βarrestin 1/2 siRNA Filamin A siRNAGRK2 siRNA 2,5 fold Expression is reduced by 7,5 fold 3 fold Beta arrestine 2 GFP ezrin Ezrin recruitment 0% 20% 40% 60% 80% 100% Control siRNA Arrestin 1+2 sirRNA FLNA sirRNAGRK2 sirRNA Ezrin
Recepteur adrénergique Beta2 AR - YFP ezrin % of recruiting colony Ezrin recruitment ControlPhentolamine 10 uM Propranolol 10 uM α-antagonist β-antagonist
Recepteur adrénergique Beta2 AR - YFP ezrin + β-agonist Ct0,00010,01110 Ezrin recruitment % of recruiting colony Isoproterenol (μM)
Recepteur adrénergique Beta2 AR - YFP ezrin β-agonist Ct0,00010,01110 Ezrin recruitment % of recruiting colony Isoproterenol (μM) +
Recepteur adrénergique Ezrin recruitment 0% 20% 40% 60% 80% 100% control Isoproterenol 1uM Isoproterenol 100nM SDF1 MIP IL8 AT2 5 à10 x Kd Ezrin recruitment 0% 20% 40% 60% 80% 100% Isoproterenol Propranolol - 10µM 10µM 1µM 1µM
Signaling des protéines G Control PKA-PS 20uM10uM Forkolin M PKA-PSForkolin M Ezrin recruitment % of recruiting colony
Activation partielle
Billes coatées, cellules hCMEC/D3 Phenylephrine α1 agoniste Isoproterenol β1 agoniste contrasteactinezrin BSA ou Metoprolol
Effet des agonistes adrénergiques Nm P-Ezrin P-Moesin Ct Isoproterenol Ezrin phosphorylation fold increase Ct Isoproterenol Ct0,00010,01110 Recrutment dEzrin % recruiting colonies Isoproterenol (μM) Ct 0,00010, Iso + Phe 10: % recruiting colonies Recrutment dEzrin Phenylephrine (μM)