Formation of toxic oligomeric alpha-synuclein species in living cells.
Formation of toxic oligomeric alpha-synuclein species in living cells.
复制标题
DOI：
10.1371/journal.pone.0001867
复制
发表时间：
2008-04-02
期刊：
影响因子：
3.7
通讯作者：
McLean PJ
中科院分区：
文献类型：
--
作者：
Outeiro TF;Putcha P;Tetzlaff JE;Spoelgen R;Koker M;Carvalho F;Hyman BT;McLean PJ
α-突触核蛋白的错误折叠、寡聚化和纤维化被认为是帕金森病（PD）和相关疾病发生和进展的中心事件。虽然纤维状α-突触核蛋白是路易体（LB）的主要成分，但最近的数据表明，前纤维状寡聚体中间体是有毒物质。然而，迄今为止，在活细胞中尚未鉴定出寡聚物种类。在这里，我们使用双分子荧光互补（BiFC）直接可视化活细胞中的α-突触核蛋白寡聚化，使我们能够研究导致α-突触核蛋白寡聚化的初始事件，聚集体形成的前体。这种新的测定为我们提供了一种工具，用于研究影响α-突触核蛋白聚集的操作如何随时间影响该过程。通过BiFC稳定α-突触核蛋白寡聚体导致细胞毒性增加，这可以在减少α-突触核蛋白寡聚体形成的过程中被Hsp 70拯救。在α-突触核蛋白中引入PD相关突变并不影响寡聚体的形成，但突变体α-突触核蛋白寡聚体的生物化学性质与野生型α-突触核蛋白的生物化学性质不同。BiFC测定法在神经退行性疾病分子基础研究中的这种新应用使得能够直接观察活细胞中的α-突触核蛋白寡聚物种类及其受Hsp 70的调节，构成了寻找突触核蛋白病治疗方法的新的重要工具。
Misfolding, oligomerization, and fibrillization of α-synuclein are thought to be central events in the onset and progression of Parkinson's disease (PD) and related disorders. Although fibrillar α-synuclein is a major component of Lewy bodies (LBs), recent data implicate prefibrillar, oligomeric intermediates as the toxic species. However, to date, oligomeric species have not been identified in living cells. Here we used bimolecular fluorescence complementation (BiFC) to directly visualize α-synuclein oligomerization in living cells, allowing us to study the initial events leading to α-synuclein oligomerization, the precursor to aggregate formation. This novel assay provides us with a tool with which to investigate how manipulations affecting α-synuclein aggregation affect the process over time. Stabilization of α-synuclein oligomers via BiFC results in increased cytotoxicity, which can be rescued by Hsp70 in a process that reduces the formation of α-synuclein oligomers. Introduction of PD-associated mutations in α-synuclein did not affect oligomer formation but the biochemical properties of the mutant α-synuclein oligomers differ from those of wild type α-synuclein. This novel application of the BiFC assay to the study of the molecular basis of neurodegenerative disorders enabled the direct visualization of α-synuclein oligomeric species in living cells and its modulation by Hsp70, constituting a novel important tool in the search for therapeutics for synucleinopathies.
登录
查看更多内容
影响因子：
56.9
作者：
Singleton, AB;Farrer, M;Gwinn-Hardy, K
通讯作者：
Gwinn-Hardy, K
DOI：
10.1073/pnas.97.2.571
发表时间：
2000-01-18
影响因子：
11.1
作者：
Conway, KA;Lee, SJ;Lansbury, PT
通讯作者：
Lansbury, PT
影响因子：
5.6
作者：
Lashuel, HA;Petre, BM;Lansbury, PT
通讯作者：
Lansbury, PT
影响因子：
48
作者：
Remy, Ingrid;Michnick, Stephen W.
通讯作者：
Michnick, Stephen W.
影响因子：
56.9
作者：
Auluck, PK;Chan, HYE;Bonini, NM
通讯作者：
Bonini, NM