The Parkinson's disease (PD) protein kinase (enzyme) LRRK2 is regulated by a chemical process called phosphorylation. To date, only a handful of the sites of phosphorylation have been investigated. These sites are important for understanding how LRRK2 is regulated and dysregulated in the altered state. We will generate immunological reagents (solutions or mixtures that cause chemical reactions) to detect altered LRRK2 when phosphorylated at these sites to determine if they are similarly dysregulated in the disease state.
We hypothesize that post-translational modification (changes in structure) of LRRK2 is a major regulatory mechanism for this disease-related kinase.
We will generate antibodies (immune system proteins) that recognize LRRK2 when phosphorylated at specific sites on the protein. Using these new tools, we will then ask if these sites are regulated by LRRK2 activity, disease mutation or in samples from individuals with PD.
Impact on Diagnosis/Treatment of Parkinson's disease:
Several sites of LRRK2 phosphorylation are currently used as markers of activity for LRRK2 inhibitor-based drug development. In several altered forms of LRRK2, phosphorylation is disrupted. By developing new agents that will detect uncharacterized phosphorylation sites, we may find new markers of activity, learn how LRRK2 is turned on or off and how this is disrupted in PD.
Next Steps for Development:
We will expand development of successful antibodies to a more specific and renewable source of reagents; these can then be validated and used in other laboratories. The kinases and phosphatases that regulate LRRK2 are new signaling hubs that may be targeted for drug development as regulators of LRRK2 function in PD. The reagents developed in this project will be crucial for identification and characterization of those kinases and phosphatases.