Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity
Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity
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DOI：
10.1073/pnas.0507360102
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发表时间：
2005-11-15
影响因子：
11.1
通讯作者：
Dawson, TM
中科院分区：
文献类型：
--
作者：
West, AB;Moore, DJ;Dawson, TM
富亮氨酸重复激酶2基因（LRRK2）突变导致迟发性帕金森病（PD），其临床表现与特发性PD难以区分。最初的研究表明，LRRK2突变是最常见的PD易感性决定因素，以常染色体显性遗传模式传播。在此，我们表征了LRRK2基因及其在人脑中的转录物，并亚克隆了显性ORF。外源表达的LRRK2蛋白迁移速度约为280 kDa，主要存在于细胞质中，但也与线粒体外膜有关。家族连锁突变G2019S或R1441C对蛋白质稳态水平、周转或定位没有明显影响。然而，使用全长重组LRRK2进行的体外激酶分析显示，自磷酸化和通用底物磷酸化的家族相关突变引起的活性增加。这些结果表明lrrk2相关疾病的功能获得机制在PD的发展中具有激酶活性的核心作用。
Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) cause late-onset Parkinson's disease (PD) with a clinical appearance indistinguishable from idiopathic PD. Initial studies suggest that LRRK2 mutations are the most common yet identified determinant of PD susceptibility, transmitted in an autosomal-dominant mode of inheritance. Herein, we characterize the LRRK2 gene and transcript in human brain and subclone the predominant ORF. Exogenously expressed LRRK2 protein migrates at approximate to 280 kDa and is present largely in the cytoplasm but also associates with the mitochondrial outer membrane. Familial-linked mutations G2019S or R1441C do not have an obvious effect on protein steady-state levels, turnover, or localization. However, in vitro kinase assays using full-length recombinant LRRK2 reveal an increase in activity caused by familial-linked mutations in both autophosphorylation and the phosphorylation of a generic substrate. These results suggest a gain-of-function mechanism for LRRK2-linked disease with a central role for kinase activity in the development of PD.