title: GTP binding is essential to the protein kinase activity of LRRK2, a causative gene product for familial Parkinson's disease.
abstract: Leucine-rich repeat kinase 2 (LRRK2), a product of a causative gene for the autosomal-dominant form of familial Parkinson's disease (PARK8), harbors a Ras-like small GTP binding protein-like (ROC) domain besides the kinase domain, although the relationship between these two functional domains remains elusive. Here we show by thin-layer chromatographic analysis that LRRK2 stably binds GTP but lacks a GTPase activity in HEK293 and Neuro-2a cells. A ROC domain mutation that converts LRRK2 to a guanine nucleotide-free form (T1348N) abolishes the kinase activity of LRRK2 as well as its phosphate incorporation upon metabolic labeling. The phosphorylation of LRRK2 was inhibited by potential inhibitors for cyclic AMP-dependent protein kinase. These data suggest that binding of GTP to the ROC domain regulates the kinase activity of LRRK2 as well as its phosphorylation by other kinase(s).
authors.affiliation: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyoku, Tokyo, 113-0033 Japan.
authors.forename: Genta
authors.initials: G
authors.lastname: Ito
authors.forename: Takuro
authors.initials: T
authors.lastname: Okai
authors.forename: Go
authors.initials: G
authors.lastname: Fujino
authors.forename: Kohsuke
authors.initials: K
authors.lastname: Takeda
authors.forename: Hidenori
authors.initials: H
authors.lastname: Ichijo
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Katada
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iwatsubo
mesh_terms: D001665:Binding Sites; D002845:Chromatography; D005192:Family Health; D020558:GTP Phosphohydrolases; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D020125:Mutation, Missense; D010300:Parkinson Disease / Q000201:enzymology*; D010766:Phosphorylation; D011485:Protein Binding; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D006160:Guanosine Triphosphate; D011494:Protein Kinases; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; D020558:GTP Phosphohydrolases