| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Valiyaveettil, Jacob Ananthan, Subramaniam Maddry, Joseph A. Gupta, Vandana Suto, Mark J. Moukha-chafiq, Omar Delucas, Lawrence J. Liu, Zhiyong Galemmo, Robert A. West, Andrew B. Volpicelli-daley, Laura A. Sen, Saurabh Pathak, Ashish K. Ross, Larry J. Kezar, Hollis White, E. Lucile Fraser, Kyle B. Moehle, Mark S. |
| Description |
Author Affiliation: Liu Z ( From the Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology and Center for Biophysical Sciences and Engineering, Department of Optometry, The University of Alabama at Birmingham, Birmingham, Alabama 35294 and.); Galemmo RA ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Fraser KB ( From the Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology and.); Moehle MS ( From the Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology and.); Sen S ( From the Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology and.); Volpicelli-Daley LA ( From the Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology and.); DeLucas LJ ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Ross LJ ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Valiyaveettil J ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Moukha-Chafiq O ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Pathak AK ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Ananthan S ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Kezar H ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); White EL ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Gupta V ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Maddry JA ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); Suto MJ ( the Drug Discovery Division, Southern Research Institute, Birmingham, Alabama 35294.); West AB ( From the Center for Neurodegeneration and Experimental Therapeutics, Department of Neurology and abwest@uab.edu.) |
| Abstract |
Pathogenic mutations in the LRRK2 gene can cause late-onset Parkinson disease. The most common mutation, G2019S, resides in the kinase domain and enhances activity. LRRK2 possesses the unique property of cis-autophosphorylation of its own GTPase domain. Because high-resolution structures of the human LRRK2 kinase domain are not available, we used novel high-throughput assays that measured both cis-autophosphorylation and trans-peptide phosphorylation to probe the ATP-binding pocket. We disclose hundreds of commercially available activity-selective LRRK2 kinase inhibitors. Some compounds inhibit cis-autophosphorylation more strongly than trans-peptide phosphorylation, and other compounds inhibit G2019S-LRRK2 more strongly than WT-LRRK2. Through exploitation of structure-activity relationships revealed through high-throughput analyses, we identified a useful probe inhibitor, SRI-29132 (11). SRI-29132 is exquisitely selective for LRRK2 kinase activity and is effective in attenuating proinflammatory responses in macrophages and rescuing neurite retraction phenotypes in neurons. Furthermore, the compound demonstrates excellent potency, is highly blood-brain barrier-permeant, but suffers from rapid first-pass metabolism. Despite the observed selectivity of SRI-29132, docking models highlighted critical interactions with residues conserved in many protein kinases, implying a unique structural configuration for the LRRK2 ATP-binding pocket. Although the human LRRK2 kinase domain is unstable and insoluble, we demonstrate that the LRRK2 homolog from ameba can be mutated to approximate some aspects of the human LRRK2 ATP-binding pocket. Our results provide a rich resource for LRRK2 small molecule inhibitor development. More broadly, our results provide a precedent for the functional interrogation of ATP-binding pockets when traditional approaches to ascertain structure prove difficult. |
| ISSN |
00219258 |
| e-ISSN |
1083351X |
| Journal |
Journal of Biological Chemistry |
| Issue Number |
47 |
| Volume Number |
289 |
| Language |
English |
| Publisher |
American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date |
2014-11-21 |
| Publisher Place |
United States |
| Access Restriction |
Open |
| Subject Keyword |
Adenosine Triphosphate Chemistry Protein Structure, Tertiary Protein-Serine-Threonine Kinases Metabolism Amino Acid Sequence Animals Binding Sites Genetics Biocatalysis Drug Effects Blotting, Western Cell Line, Tumor Cells, Cultured Hep G2 Cells Kinetics Mice Models, Molecular Molecular Sequence Data Molecular Structure Mutation Phosphorylation Protein Binding Protein Kinase Inhibitors Pharmacology Pyridazines Sequence Homology, Amino Acid Small Molecule Libraries Structure-Activity Relationship Triazoles Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Biochemistry Molecular Biology |
| Content Type |
Text |
| Resource Type |
Article |
| Subject |
Cell Biology Biochemistry Molecular Biology |
