title: Leucine-rich repeat kinase 2 associates with lipid rafts.
abstract: Leucine-Rich Repeat Kinase 2 (LRRK2) is a causative gene for the autosomal dominant form of Parkinson's disease (PD). The gene encodes the approximately 280 kDa LRRK2 protein composed of domains such as leucine-rich repeats, Ras in complex proteins (Roc) followed by C-terminal of Roc (COR), mitogen-activated protein kinase kinase kinase (MAPKKK) and WD40. However, the normal function of the protein as well as its contribution to the pathogenesis of PD remains largely unknown. Here we describe the localization of LRRK2 in Golgi apparatus, plasma membrane and synaptic vesicles in cultured cells including mouse primary neurons. The membrane association of LRRK2 resists solubilization by ice-cold 1% Triton X-100, indicating its association through lipid rafts. To investigate whether mutations found in PD patients affect the localization of LRRK2, we transfected various LRRK2 mutants into cultured cells and performed fractionation experiments. Unexpectedly, the mutants are collected in both membrane and soluble fractions in a manner similar to wild type (WT). I2020T mutant LRRK2 associates with lipid rafts, similar to the WT. The lipid raft association of LRRK2 mutants as well as WT LRRK2 suggests that alteration of LRRK2 function on lipid rafts contributes to the pathogenesis of PD.
authors.affiliation: Department of Neurology, Juntendo University School of Medicine, Bunkyo, Tokyo, Japan.
authors.forename: Taku
authors.initials: T
authors.lastname: Hatano
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Kubo
authors.forename: Satoshi
authors.initials: S
authors.lastname: Imai
authors.forename: Masahiro
authors.initials: M
authors.lastname: Maeda
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Ishikawa
authors.forename: Yoshikuni
authors.initials: Y
authors.lastname: Mizuno
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Hattori
mesh_terms: D000818:Animals; D000906:Antibodies; D001921:Brain / Q000201:enzymology; D019556:COS Cells; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D022162:Cytoplasmic Vesicles / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D021962:Membrane Microdomains / Q000201:enzymology*; D051379:Mice; D009154:Mutation; D009474:Neurons / Q000201:enzymology; D010300:Parkinson Disease / Q000209:etiology; D017729:Presynaptic Terminals / Q000201:enzymology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D014162:Transfection; D021601:trans-Golgi Network / Q000201:enzymology
chemical_list: D000906:Antibodies; D034741:RNA, Small Interfering; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; C495279:Lrrk2 protein, mouse; D017346:Protein Serine-Threonine Kinases