The LRRK2 G2385R variant is a partial loss-of-function mutation that affects synaptic vesicle trafficking through altered protein interactions.
The LRRK2 G2385R variant is a partial loss-of-function mutation that affects synaptic vesicle trafficking through altered protein interactions.
复制标题
DOI：
10.1038/s41598-017-05760-9
复制
发表时间：
2017-07-14
影响因子：
4.6
通讯作者：
Piccoli G
中科院分区：
文献类型：
--
作者：
Carrion MDP;Marsicano S;Daniele F;Marte A;Pischedda F;Di Cairano E;Piovesana E;von Zweydorf F;Kremmer E;Gloeckner CJ;Onofri F;Perego C;Piccoli G
富含亮氨酸重复序列激酶2基因（LRRK 2）突变与家族性帕金森病（PD）相关。LRRK 2蛋白含有几个功能结构域，包括在其N-和C-末端的蛋白质-蛋白质相互作用结构域。在这项研究中，我们分析了LRRK 2的N-和C-末端结构域的功能特点。我们结合TIRF显微镜和synaptopHluorin检测来观察突触囊泡的运输。我们发现，N-和C-末端结构域对突触囊泡动力学有相反的影响。生物化学分析表明，不同的蛋白质结合在两个末端，即N端部分的β3-Cav2.1和C端结构域的β-Actin和Synapsin I。在C-末端WD 40结构域内的序列变体（G2385 R）增加了PD的风险。互补的生物化学和成像方法显示，G2385 R变体改变了LRRK 2相互作用的强度和质量，并增加了突触囊泡的融合。我们的数据表明，G2385 R变体的行为就像一个功能丧失突变，模仿活动驱动的事件。突变体LRRK 2的支架能力受损，导致受干扰的囊泡运输，可能会出现作为一个共同的病理生理学分母，通过不同的LRRK 2病理突变引起疾病。
Mutations in the Leucine-rich repeat kinase 2 gene (LRRK2) are associated with familial Parkinson’s disease (PD). LRRK2 protein contains several functional domains, including protein-protein interaction domains at its N- and C-termini. In this study, we analyzed the functional features attributed to LRRK2 by its N- and C-terminal domains. We combined TIRF microscopy and synaptopHluorin assay to visualize synaptic vesicle trafficking. We found that N- and C-terminal domains have opposite impact on synaptic vesicle dynamics. Biochemical analysis demonstrated that different proteins are bound at the two extremities, namely β3-Cav2.1 at N-terminus part and β-Actin and Synapsin I at C-terminus domain. A sequence variant (G2385R) harboured within the C-terminal WD40 domain increases the risk for PD. Complementary biochemical and imaging approaches revealed that the G2385R variant alters strength and quality of LRRK2 interactions and increases fusion of synaptic vesicles. Our data suggest that the G2385R variant behaves like a loss-of-function mutation that mimics activity-driven events. Impaired scaffolding capabilities of mutant LRRK2 resulting in perturbed vesicular trafficking may arise as a common pathophysiological denominator through which different LRRK2 pathological mutations cause disease.
登录
查看更多内容
影响因子：
15.1
作者：
Belluzzi E;Gonnelli A;Cirnaru MD;Marte A;Plotegher N;Russo I;Civiero L;Cogo S;Carrion MP;Franchin C;Arrigoni G;Beltramini M;Bubacco L;Onofri F;Piccoli G;Greggio E
通讯作者：
Greggio E
DOI：
10.1074/mcp.m114.041012
发表时间：
2015-01
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Keilhauer EC;Hein MY;Mann M
通讯作者：
Mann M
影响因子：
4.8
作者：
Greggio, Elisa;Zambrano, Ibardo;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
影响因子：
5.1
作者：
Bauer, M.;Kinkl, N.;Ueffing, M.
通讯作者：
Ueffing, M.
影响因子：
5.3
作者：
Beccano-Kelly DA;Kuhlmann N;Tatarnikov I;Volta M;Munsie LN;Chou P;Cao LP;Han H;Tapia L;Farrer MJ;Milnerwood AJ
通讯作者：
Milnerwood AJ