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Fluctuation Imaging of LRRK2 Reveals that the G2019S Mutation Alters Spatial and Membrane Dynamics
- Source :
- Molecules, Volume 25, Issue 11, Molecules, Vol 25, Iss 2561, p 2561 (2020)
- Publication Year :
- 2020
-
Abstract
- Mutations within the Leucine-Rich Repeat Kinase 2 (LRRK2) gene are the most common genetic cause of autosomal and sporadic Parkinson&rsquo<br />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<br />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 with a reduction in transferrin endocytosis in cells expressing G2019S, which indicates disruption of endocytic protein recruitment near the plasma membrane. Overall, this study uncovered multiple dynamic alterations to the LRRK2 protein as a result of the G2019S mutation&mdash<br />all of which could lead to neurodegeneration associated with PD.
- Subjects :
- Endocytic cycle
Pharmaceutical Science
Analytical Chemistry
fluorescence fluctuation spectroscopy
0302 clinical medicine
Cytosol
Drug Discovery
Fluorescence Resonance Energy Transfer
Phosphorylation
0303 health sciences
education.field_of_study
Microscopy, Confocal
biology
Chemistry
Vesicle
Transferrin
Parkinson Disease
LRRK2
FRET-Phasor
Endocytosis
Cell biology
Chemistry (miscellaneous)
Molecular Medicine
FLIM
Population
Green Fluorescent Proteins
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Clathrin
Article
lcsh:QD241-441
03 medical and health sciences
lcsh:Organic chemistry
Protein Domains
Cell Line, Tumor
Humans
Physical and Theoretical Chemistry
education
030304 developmental biology
Adaptor Proteins, Signal Transducing
Organic Chemistry
Cell Membrane
nervous system diseases
Membrane protein
Mutation
biology.protein
Parkinson’s disease
Rab
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 14203049
- Volume :
- 25
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- Molecules (Basel, Switzerland)
- Accession number :
- edsair.doi.dedup.....a49c1e5d34223c9c7151a292f1f2dcbf