title: Parkinson disease-associated mutation R1441H in LRRK2 prolongs the "active state" of its GTPase domain.
abstract: Mutation in leucine-rich-repeat kinase 2 (LRRK2) is a common cause of Parkinson disease (PD). A disease-causing point mutation R1441H/G/C in the GTPase domain of LRRK2 leads to overactivation of its kinase domain. However, the mechanism by which this mutation alters the normal function of its GTPase domain [Ras of complex proteins (Roc)] remains unclear. Here, we report the effects of R1441H mutation (RocR1441H) on the structure and activity of Roc. We show that Roc forms a stable monomeric conformation in solution that is catalytically active, thus demonstrating that LRRK2 is a bona fide self-contained GTPase. We further show that the R1441H mutation causes a twofold reduction in GTPase activity without affecting the structure, thermal stability, and GDP-binding affinity of Roc. However, the mutation causes a twofold increase in GTP-binding affinity of Roc, thus suggesting that the PD-causing mutation R1441H traps Roc in a more persistently activated state by increasing its affinity for GTP and, at the same time, compromising its GTP hydrolysis.
authors.affiliation: Department of Biochemistry and Molecular Biology and Stark Neurosciences Institute, Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Jingling
authors.initials: J
authors.lastname: Liao
authors.forename: Chun-Xiang
authors.initials: CX
authors.lastname: Wu
authors.forename: Christopher
authors.initials: C
authors.lastname: Burlak
authors.forename: Sheng
authors.initials: S
authors.lastname: Zhang
authors.forename: Heather
authors.initials: H
authors.lastname: Sahm
authors.forename: Mu
authors.initials: M
authors.lastname: Wang
authors.forename: Zhong-Yin
authors.initials: ZY
authors.lastname: Zhang
authors.forename: Kurt W
authors.initials: KW
authors.lastname: Vogel
authors.forename: Mark
authors.initials: M
authors.lastname: Federici
authors.forename: Steve M
authors.initials: SM
authors.lastname: Riddle
authors.forename: R Jeremy
authors.initials: RJ
authors.lastname: Nichols
authors.forename: Dali
authors.initials: D
authors.lastname: Liu
authors.forename: Mark R
authors.initials: MR
authors.lastname: Cookson
authors.forename: Todd A
authors.initials: TA
authors.lastname: Stone
authors.forename: Quyen Q
authors.initials: QQ
authors.lastname: Hoang
mesh_terms: D015153:Blotting, Western; D002850:Chromatography, Gel; D002942:Circular Dichroism; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D013058:Mass Spectrometry; D008958:Models, Molecular; D020125:Mutation, Missense / Q000235:genetics*; D010300:Parkinson Disease / Q000235:genetics*; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; D020558:GTP Phosphohydrolases
keywords: dimer; monomer; neurodegenerative disease; oligomeric states