Effect of 4E-BP and eIF4E complex on phosphorylation of 4E-BP by LRRK2 in vitro.

(A) Time-course assays were performed for LRRK2 wild-type (upper panel) by measuring the incorporation of [γ³³P] ATP at different time points (0, 5, 15, 30 and 60 mins) and at 3 µM concentrations of 4E-BP (upper left panel), 3 µM concentrations of eIF4E (upper middle panel) and 3 µM concentrations of 4E-BP and 3 µM concentrations of eIF4E (upper right panel), respectively, followed by SDS-PAGE and autoradiography. (B) Phosphorylation of 4E-BP by LRRK2-wild type were plotted as a function of time.