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Updated: Mar 17, 2026
Assaying the Kinase Activity of LRRK2 in vitro
Published on: January 18, 2012
Functional properties of LRRK2 mutations in Taiwanese Parkinson disease
Kuo-Hsuan Chang1, Chiung-Mei Chen1, Chih-Hsin Lin1
1Department of Neurology, Chang Gung Memorial Hospital-Linkou Medical Center, Chang Gung University College of Medicine, Taipei 10507, Taiwan.
Background/Purpose:
Leucine-rich repeat kinase 2 (LRRK2) is a large protein encoding multiple functional domains. Mutations within different LRRK2 domains have been considered to be involved in the development of Parkinson disease by different mechanisms. Our previous study found three LRRK2 mutations-p.R767H, p.S885N, and p.R1441H-in Taiwanese patients with Parkinson disease.
Methods:
We evaluated the functional properties of LRRK2 p.R767H, p.S885N, and p.R1441H mutations by overexpressing them in human embryonic kidney 293 and neuroblastoma SK-N-SH cells. The common p.G2019S mutation in the kinase domain was included for comparison.
Results:
In 293 cells, overexpressed p.R1441H-but not p.R767H, p.S885N, or p.G2019-increased GTP binding affinity to prolong the active state. Overexpressed p.R1441H and p.G2019S generated inclusions in 293 cells. In SK-N-SH cells, the α-synuclein was coexpressed with wild type as well as mutated p.R767H, p.S885N, p.R1441H, and p.G2019 LRRK2 proteins. Part of the perinuclear inclusions formed by p.R1441H and p.G2019S were colocalized with α-synuclein. Additionally, p.S885N and p.R1441H mutations caused reduced interaction between LRRK2 and ARHGEF7, a putative guanine nucleotide exchange factor for LRRK2, whereas this interaction was well preserved in p.R767H and p.G2019S mutations.
Conclusion:
Our study suggests that p.R1441H protein facilitates the formation of intracellular inclusions, compromises GTP hydrolysis by increasing its affinity for GTP, and reduces its interaction with ARHGEF7.
Insights
The R1441H mutation in Leucine-rich repeat kinase 2 (LRRK2) protein promotes Parkinson disease pathology by forming inclusions and impairing GTP hydrolysis. This LRRK2 mutation also reduces interaction with ARHGEF7.
Area of Science:
- Neuroscience
- Genetics
- Molecular Biology
Background:
- Leucine-rich repeat kinase 2 (LRRK2) is implicated in Parkinson disease pathogenesis.
- LRRK2 mutations affect different functional domains, potentially through various mechanisms.
- Three specific LRRK2 mutations (p.R767H, p.S885N, p.R1441H) were identified in Taiwanese Parkinson disease patients.
Purpose of the Study:
- To investigate the functional consequences of LRRK2 mutations p.R767H, p.S885N, and p.R1441H.
- To compare these mutations with the common LRRK2 p.G2019S mutation.
Main Methods:
- Overexpression of wild-type and mutant LRRK2 proteins (p.R767H, p.S885N, p.R1441H, p.G2019S) in human embryonic kidney 293 and neuroblastoma SK-N-SH cells.
- Assessment of GTP binding affinity and formation of intracellular inclusions.
- Evaluation of co-expression with α-synuclein and interaction with ARHGEF7.
Main Results:
- The p.R1441H mutation significantly increased GTP binding affinity, prolonging LRRK2's active state.
- Overexpressed p.R1441H and p.G2019S LRRK2 proteins formed inclusions in 293 cells, with p.R1441H inclusions colocalizing with α-synuclein in SK-N-SH cells.
- p.S885N and p.R1441H mutations reduced the interaction between LRRK2 and ARHGEF7, unlike p.R767H and p.G2019S.
Conclusions:
- The p.R1441H LRRK2 mutation contributes to Parkinson disease by promoting intracellular inclusion formation.
- This mutation impairs GTP hydrolysis, leading to a prolonged active state of LRRK2.
- Reduced interaction between LRRK2 and ARHGEF7 is a consequence of the p.R1441H and p.S885N mutations.
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