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Updated: Dec 22, 2025
Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
Published on: December 14, 2017
LRRK2 Is Recruited to Phagosomes and Co-recruits RAB8 and RAB10 in Human Pluripotent Stem Cell-Derived Macrophages
Heyne Lee1, Rowan Flynn1, Ishta Sharma1
1Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
Abstract:
The Parkinson's disease-associated gene, LRRK2, is also associated with immune disorders and infectious disease and is expressed in immune subsets. Here, we characterize a platform for interrogating the expression and function of endogenous LRRK2 in authentic human phagocytes using human induced pluripotent stem cell-derived macrophages and microglia. Endogenous LRRK2 is expressed and upregulated by interferon-γ in these cells, including a 187-kDa cleavage product. Using LRRK2 knockout and G2019S isogenic repair lines, we find that LRRK2 is not involved in initial phagocytic uptake of bioparticles but is recruited to LAMP1+/RAB9+ "maturing" phagosomes, and LRRK2 kinase inhibition enhances its residency at the phagosome. Importantly, LRRK2 is required for RAB8a and RAB10 recruitment to phagosomes, implying that LRRK2 operates at the intersection between phagosome maturation and recycling pathways in these professional phagocytes.
Insights
The Parkinson's disease gene LRRK2 is expressed in human immune cells and influences phagosome maturation. LRRK2 kinase inhibition affects its localization and recruitment of other proteins to phagosomes.
Area of Science:
- Immunology
- Cell Biology
- Neuroscience
Background:
- The leucine-rich repeat kinase 2 (LRRK2) gene is linked to Parkinson's disease and immune disorders.
- LRRK2 is expressed in various immune cells, suggesting a role in immune function.
- Understanding LRRK2 function in professional phagocytes is crucial for its role in disease.
Purpose of the Study:
- To characterize the expression and function of endogenous LRRK2 in human phagocytes.
- To investigate LRRK2's role in phagosome maturation and recycling pathways.
- To explore the impact of LRRK2 kinase activity on its phagosomal localization.
Main Methods:
- Utilized human induced pluripotent stem cell-derived macrophages and microglia.
- Employed LRRK2 knockout and G2019S isogenic repair cell lines.
- Analyzed LRRK2 expression, localization, and recruitment of RAB GTPases to phagosomes.
Main Results:
- Endogenous LRRK2 is expressed and upregulated by interferon-γ in macrophages and microglia, with a notable 187-kDa cleavage product.
- LRRK2 is not essential for initial phagocytic uptake but is recruited to maturing phagosomes (LAMP1+/RAB9+).
- LRRK2 kinase inhibition increases its residence time at the phagosome and is critical for RAB8a and RAB10 recruitment.
Conclusions:
- LRRK2 plays a significant role in the maturation and recycling of phagosomes in human phagocytes.
- LRRK2 kinase activity regulates its localization and interaction with key trafficking proteins.
- These findings highlight LRRK2 as a potential therapeutic target at the crossroads of immune cell function and neurodegenerative disease.
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