Membrane localization of LRRK2 is associated with increased formation of the highly active LRRK2 dimer and changes in its phosphorylation.
Membrane localization of LRRK2 is associated with increased formation of the highly active LRRK2 dimer and changes in its phosphorylation.
复制标题
DOI：
10.1021/bi100157u
复制
发表时间：
2010-07-06
期刊：
影响因子：
2.9
通讯作者：
LaVoie, Matthew J.
中科院分区：
文献类型：
--
作者：
Berger, Zdenek;Smith, Kelsey A.;LaVoie, Matthew J.
富含亮氨酸重复序列激酶2（LRRK 2）的常染色体显性突变是帕金森病（PD）最常见的遗传原因。尽管存在多个结构域，但LRRK 2的激酶活性被认为代表了蛋白质的主要功能。LRRK 2激酶活性的改变被认为是其PD连锁突变的发病机制的基础;然而，关于LRRK 2功能的基本方面的许多问题仍然不清楚，包括LRRK 2调节的细胞机制及其在细胞内独特分布的重要性。在这里，我们第一次证明了野生型LRRK 2的亚细胞定位与四种不同的生化特性的变化有关，这四种生化特性可能对LRRK 2的功能至关重要。我们的数据首次表明，野生型LRRK 2二聚体具有更大的激酶活性比其更丰富的单体对应物。重要的是，我们表明，这种活化形式的LRRK 2基本上富集在表达内源性或外源性LRRK 2的细胞膜上，并且LRRK 2的膜相关部分同样具有比胞质LRRK 2更大的激酶活性。此外，膜相关LRRK 2比胞质LRRK 2更有效地结合GTP，但表现出较低程度的磷酸化。我们的观察结果表明，多种事件，包括改变蛋白质-蛋白质相互作用和翻译后修饰，有助于调节LRRK 2的功能，通过调节膜协会和复杂的装配。这些发现可能对细胞内LRRK 2功能的位点、真正LRRK 2底物的鉴定和定位以及设计LRRK 2小分子抑制剂的努力具有影响。
Autosomal dominant mutations in the leucine rich repeat kinase 2 (LRRK2) are the most common genetic cause of Parkinson’s disease (PD). Despite the presence of multiple domains, the kinase activity of LRRK2 is thought to represent the primary function of the protein. Alterations in LRRK2 kinase activity are thought to underlie the pathogenesis of its PD-linked mutations; however, many questions regarding basic aspects of LRRK2 function remain unclear, including the cellular mechanisms of LRRK2 regulation and the importance of its unique distribution within the cell. Here, we demonstrate for the first time that the subcellular localization of wild-type LRRK2 is associated with changes in four distinct biochemical properties likely crucial for LRRK2 function. Our data demonstrate for the first time that the wild-type LRRK2 dimer possesses greater kinase activity than its more abundant monomeric counterpart. Importantly, we show that this activated form of LRRK2 is substantially enriched at the membrane of cells expressing endogenous or exogenous LRRK2, and that the membrane-associated fraction of LRRK2 likewise possesses greater kinase activity than cytosolic LRRK2. In addition, membrane-associated LRRK2 binds GTP more efficiently than cytosolic LRRK2, but demonstrates a lower degree of phosphorylation. Our observations suggest that multiple events, including altered protein-protein interactions and post-translational modification, contribute to the regulation of LRRK2 function, through modulating membrane association and complex assembly. These findings may have implications for the sites of LRRK2 function within the cell, the identification and localization of bona fide LRRK2 substrates, and efforts to design small molecule inhibitors of LRRK2.
登录
查看更多内容
DOI：
10.1016/j.bbrc.2009.08.163
发表时间：
2009-11-20
影响因子：
3.1
作者：
Greggio, Elisa;Taymans, Jean-Marc;Zhen, Eugene Yuejun;Ryder, John;Vancraenenbroeck, Renee;Beilina, Alexandra;Sun, Peng;Deng, Junpeng;Jaffe, Howard;Baekelandt, Veerle;Merchant, Kalpana;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
影响因子：
11.2
作者：
Biskup, Saskia;Moore, Darren J.;Dawson, Valina L.
通讯作者：
Dawson, Valina L.
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：
2.9
作者：
Ito, Genta;Okai, Takuro;Iwatsubo, Takeshi
通讯作者：
Iwatsubo, Takeshi
影响因子：
3.7
作者：
Guo, Luxuan;Gandhi, Payal N.;Chen, Shu G.
通讯作者：
Chen, Shu G.