Molecules, Vol. 25, Pages 2561: Fluctuation Imaging of LRRK2 Reveals That the G2019S Mutation Alters Spatial and Membrane Dynamics

Molecules, Vol. 25, Pages 2561: Fluctuation Imaging of LRRK2 Reveals That the G2019S Mutation Alters Spatial and Membrane Dynamics Molecules doi: 10.3390/molecules25112561 Authors: Sanstrum Goo Holden Delgado Nguyen Lee James Mutations within the Leucine-Rich Repeat Kinase 2 (LRRK2) gene are the most common genetic cause of autosomal and sporadic Parkinson’s disease (PD). LRRK2 is a large multidomain kinase that has reported interactions with several membrane proteins, including Rab and Endophilin, and has recently been proposed to function as a regulator of vesicular trafficking. It is unclear whether or how the spatiotemporal organization of the protein is altered due to LRRK2 activity. Therefore, we utilized fluctuation-based microscopy along with FLIM/FRET to examine the cellular properties and membrane recruitment of WT LRRK2-GFP (WT) and the PD mutant G2019S LRRK2-GFP (G2019S). We show that both variants can be separated into two distinct populations within the cytosol; a freely diffusing population associated with monomer/dimer species and a slower, likely vesicle-bound population. G2019S shows a significantly higher propensity to self-associate in both the cytosol and membrane regions when compared to WT. G2019S expression also resulted in increased hetero-interactions with Endophilin A1 (EndoA1), reduced cellular vesicles, and altered clathrin puncta dynamics associated with the plasma membrane. This finding was associated wit...
Source: Molecules - Category: Chemistry Authors: Tags: Article Source Type: research