Fluorescence Fluctuation Spectroscopy Assays to Monitor the Activity of LRRK2
LRRK2 Pharmacodynamic Assays, 2013
Objective/Rationale: † † † † † ††
LRRK2 is a major protein target for therapeutic treatment of Parkinsonís disease (PD). The lack of assays (experiment set-ups) available for measuring the activity of LRRK2 has hindered the progress of a LRRK2-targeted PD treatment. We will develop a fluorescence-based assay(s) to study LRRK2 activity in live cells. The successful development of these assays will allow rapid screening for new potential LRRK2 therapeutic compounds.
Project Description: † † † † † ††
The purpose of this study is to measure the light emission properties of fluorescently tagged LRRK2 in live cells and correlate this emission to LRRK2 activity. This will be accomplished by creating a general outline of LRRK2 activity based on its properties using the fluorescence from cells containing wild-type (not mutated) LRRK2, the Parkinsonís disease form G2019S and an inactive form. From this, we will test our outline of activity by measuring the emission properties of fluorescently tagged LRRK2 while in the presence of compounds that inhibit its activity. This analysis should reveal a common signal to follow that will allow for determination of LRRK2 activity in cells.
Relevance to Diagnosis/Treatment of Parkinsonís Disease: † † † † † † † † † ††
These assays may have a significant impact on the advancement on our understanding of LRRK2 biological function and development of drugs aimed at the LRRK2 activity.†
Anticipated Outcome: † † † † †
We expect to develop a functional assay that allows for quantitatively assessing the activity of LRRK2 in cells. This type of assay is advantageous as it monitors the activity over a rapid time scale and provides researchers the ability to modify LRRK2 activity conditions to watch this change in real time.
Assistant Researcher at University of Hawaii at Manoa
Location: Honolulu, Hawaii, United States