{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T19:36:35Z","timestamp":1781897795862,"version":"3.54.5"},"reference-count":40,"publisher":"Portland Press Ltd.","issue":"1","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2009,11,15]]},"abstract":"<jats:p>The LRRK2 (leucine-rich repeat protein kinase-2) is mutated in a significant number of Parkinson's disease patients, but little is known about its regulation and function. A common mutation changing Gly2019 to serine enhances catalytic activity, suggesting that small-molecule inhibitors might have utility in treating Parkinson's disease. We employed various approaches to explore the substrate-specificity requirements of LRRK2 and elaborated a peptide substrate termed Nictide, that had 20-fold lower Km and nearly 2-fold higher Vmax than the widely deployed LRRKtide substrate. We demonstrate that LRRK2 has marked preference for phosphorylating threonine over serine. We also observed that several ROCK (Rho kinase) inhibitors such as Y-27632 and H-1152, suppressed LRRK2 with similar potency to which they inhibited ROCK2. In contrast, GSK429286A, a selective ROCK inhibitor, did not significantly inhibit LRRK2. We also identified a mutant LRRK2[A2016T] that was normally active, but resistant to H-1152 and Y-27632, as well as sunitinib, a structurally unrelated multikinase inhibitor that, in contrast with other compounds, suppresses LRRK2, but not ROCK. We have also developed the first sensitive antibody that enables measurement of endogenous LRRK2 protein levels and kinase activity as well as shRNA (short hairpin RNA) methods to reduce LRRK2 expression. Finally, we describe a pharmacological approach to validate whether substrates are phosphorylated by LRRK2 and use this to provide evidence that LRRK2 may not be rate-limiting for the phosphorylation of the proposed substrate moesin. The findings of the present study will aid with the investigation of LRRK2.<\/jats:p>","DOI":"10.1042\/bj20091035","type":"journal-article","created":{"date-parts":[[2009,9,10]],"date-time":"2009-09-10T13:39:53Z","timestamp":1252589993000},"page":"47-60","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":184,"title":["Substrate specificity and inhibitors of LRRK2, a protein kinase mutated in Parkinson's disease"],"prefix":"10.1042","volume":"424","author":[{"given":"R.\u00a0Jeremy","family":"Nichols","sequence":"first","affiliation":[{"name":"MRC Protein Phosphorylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Dzamko","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica\u00a0E.","family":"Hutti","sequence":"additional","affiliation":[{"name":"Department of Systems Biology, Harvard Medical School, Boston, MA 02115, U.S.A."},{"name":"Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, MA 02215, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lewis\u00a0C.","family":"Cantley","sequence":"additional","affiliation":[{"name":"Department of Systems Biology, Harvard Medical School, Boston, MA 02115, U.S.A."},{"name":"Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, MA 02215, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Deak","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Moran","sequence":"additional","affiliation":[{"name":"Division of Signal Transduction Therapy Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Bamborough","sequence":"additional","affiliation":[{"name":"Molecular Discovery Research, GlaxoSmithKline Pharmaceuticals R&D, Medicines Research Centre, Gunnels Wood Road, Stevenage, Herts. 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