title: Ser1292 autophosphorylation is an indicator of LRRK2 kinase activity and contributes to the cellular effects of PD mutations.
abstract: Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common cause of familial Parkinson's disease (PD). Although biochemical studies have shown that certain PD mutations confer elevated kinase activity in vitro on LRRK2, there are no methods available to directly monitor LRRK2 kinase activity in vivo. We demonstrate that LRRK2 autophosphorylation on Ser(1292) occurs in vivo and is enhanced by several familial PD mutations including N1437H, R1441G/C, G2019S, and I2020T. Combining two PD mutations together further increases Ser(1292) autophosphorylation. Mutation of Ser(1292) to alanine (S1292A) ameliorates the effects of LRRK2 PD mutations on neurite outgrowth in cultured rat embryonic primary neurons. Using cell-based and pharmacodynamic assays with phosphorylated Ser(1292) as the readout, we developed a brain-penetrating LRRK2 kinase inhibitor that blocks Ser(1292) autophosphorylation in vivo and attenuates the cellular consequences of LRRK2 PD mutations in vitro. These data suggest that Ser(1292) autophosphorylation may be a useful indicator of LRRK2 kinase activity in vivo and may contribute to the cellular effects of certain PD mutations.
authors.affiliation: Department of Neurosciences, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Zejuan
authors.initials: Z
authors.lastname: Sheng
authors.forename: Shuo
authors.initials: S
authors.lastname: Zhang
authors.forename: Daisy
authors.initials: D
authors.lastname: Bustos
authors.forename: Tracy
authors.initials: T
authors.lastname: Kleinheinz
authors.forename: Claire E
authors.initials: CE
authors.lastname: Le Pichon
authors.forename: Sara L
authors.initials: SL
authors.lastname: Dominguez
authors.forename: Hilda O
authors.initials: HO
authors.lastname: Solanoy
authors.forename: Jason
authors.initials: J
authors.lastname: Drummond
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Zhang
authors.forename: Xiao
authors.initials: X
authors.lastname: Ding
authors.forename: Fang
authors.initials: F
authors.lastname: Cai
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Song
authors.forename: Xianting
authors.initials: X
authors.lastname: Li
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Yue
authors.forename: Marcel P
authors.initials: MP
authors.lastname: van der Brug
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Burdick
authors.forename: Janet
authors.initials: J
authors.lastname: Gunzner-Toste
authors.forename: Huifen
authors.initials: H
authors.lastname: Chen
authors.forename: Xingrong
authors.initials: X
authors.lastname: Liu
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Estrada
authors.forename: Zachary K
authors.initials: ZK
authors.lastname: Sweeney
authors.forename: Kimberly
authors.initials: K
authors.lastname: Scearce-Levie
authors.forename: John G
authors.initials: JG
authors.lastname: Moffat
authors.forename: Donald S
authors.initials: DS
authors.lastname: Kirkpatrick
authors.forename: Haitao
authors.initials: H
authors.lastname: Zhu
mesh_terms: D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D006160:Guanosine Triphosphate / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D010300:Parkinson Disease / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D010766:Phosphorylation / Q000187:drug effects; D010768:Phosphoserine / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats
chemical_list: D050505:Mutant Proteins; D047428:Protein Kinase Inhibitors; D010768:Phosphoserine; D006160:Guanosine Triphosphate; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; C495279:Lrrk2 protein, mouse; D017346:Protein Serine-Threonine Kinases