High-Throughput Cell-Based Assay for Lrrk2 Activity

Reference #:

2007-327

Inventors/Contributors

Matthew J. Farrer, Ph.D., Justus Daechsel, Ph.D.

Description

Leucine-rich repeat kinase 2 (lrrk2) exits as a dimer, and with associated proteins probably forms a higher molecular weight multimer that requires GTP/GDP binding and phosphorylation for activation of its intrinsic serine-threonine kinase (Daechsel et al., 2006). Based on the position of pathogenic amino acid substitutions within the protein, we have devised a parsimonius model that now explains lrrk2's function and suggests several mechanisms whereby small molecule activators and/or inhibitors of lrrk2 intrinsic kinase activity can be modulated. As leucocytes express large amounts of lrrk2 protein, and lrrk2 R1441C, Y1699C and G2019S patient leucocytes and EBV-transformed lymphoblastoid cell lines are readily available, we describe a high-throughput cell-based assay to monitor lrrk2 activity by assessing downstream targets, including Hsp27 and moesin (and phosphorylated forms), within an appropriate physiologic context.

Patent Status

None