LRRK2 regulates synaptic vesicle endocytosis
LRRK2 regulates synaptic vesicle endocytosis
复制标题
DOI：
10.1016/j.yexcr.2008.02.015
复制
发表时间：
2008-06-10
影响因子：
3.7
通讯作者：
Seol, Wongi
中科院分区：
文献类型：
--
作者：
Shin, Narae;Jeong, Hyerhan;Seol, Wongi
富含亮氨酸的重复激酶 2 (LRRK2) 已被确定为 PARK8 位点的缺陷基因，可导致常染色体显性帕金森病 (PD)。尽管在家族性和散发性PD患者中发现了多种LRRK2突变，但其生理功能尚不清楚。在这项研究中，我们利用酵母双杂交筛选，报告了 Rab5b 被鉴定为 LRRK2 相互作用蛋白。事实上，我们的 GST 下拉和免疫共沉淀测定表明它与 LRRK2 特异性相互作用。此外，亚细胞分级分离和免疫细胞化学分析证实，两种蛋白质的一部分共定位于突触小泡中。有趣的是，我们发现通过原代神经元细胞中内源性 LRRK2 的过表达或敲低来改变 LRRK2 表达会显着损害突触小泡的内吞作用。此外，这种内吞作用缺陷可以通过功能性 Rab5b 蛋白的共表达来弥补，但不能通过其非活性形式来弥补。综上所述，我们认为 LRRK2 连同其与 Rab5b 的相互作用，通过调节突触小泡的内吞作用在突触功能中发挥重要作用。 (c) 2008 Elsevier Inc. 保留所有权利。
The leucine-rich repeat kinase 2 (LRRK2) has been identified as the defective gene at the PARK8 locus causing the autosomal dominant form of Parkinson's disease (PD). Although several LRRK2 mutations were found in familial as well as sporadic PD patients, its physiological functions are not clearly defined. In this study, using yeast two-hybrid screening, we report the identification of Rab5b as an LRRK2-interacting protein. Indeed, our GST pull down and co-immunoprecipitation assays showed that it specifically interacts with LRRK2. In addition, subcellular fractionation and immunocytochemical analyses confirmed that a fraction of both proteins co-localize in synaptic vesicles. Interestingly, we found that alteration of LRRK2 expression by either overexpression or knockdown of endogenous LRRK2 in primary neuronal cells significantly impairs synaptic vesicle endocytosis. Furthermore, this endocytosis defect was rescued by co-expression of functional Rab5b protein, but not by its inactive form. Taken together, we propose that LRRK2, in conjunction with its interaction with Rab5b, plays an important role in synaptic function by modulating the endocytosis of synaptic vesicles. (c) 2008 Elsevier Inc. All rights reserved.