Familial PD–associated and functional mutations in LRRK2 modulate the interaction with ArfGAP1.

<p>(A) Differential interaction of YFP-tagged ArfGAP1 with WT and PD-associated mutant forms of FLAG-tagged human LRRK2 following IP with anti-FLAG antibody from HEK-293T cells. Densitometric analysis reveals a non-significant increase in the interaction of ArfGAP1 with R1441C, Y1699C and G2019S LRRK2 compared to WT LRRK2. (B) Functional GTPase mutations in FLAG-LRRK2 impair the interaction with ArfGAP1-YFP following IP with anti-FLAG antibody from HEK-293T cells. Densitometry reveals a significant reduction of the interaction of ArfGAP1 with T1348N and R1398L LRRK2 compared to WT LRRK2. Data represent the level of interaction of ArfGAP1 with LRRK2 expressed as a percent of the interaction with WT LRRK2. The levels of ArfGAP1 IP were first normalized to ArfGAP1 input levels, and then further normalized to LRRK2 IP levels. Bars represent the mean ± SEM (<i>n</i> = 3 experiments). *<i>P</i><0.05 compared to WT LRRK2 by one-way ANOVA with Newman-Keuls post-hoc analysis. <i>ns</i>, non-significant.