LRRK2 phosphorylates pre-synaptic N-ethylmaleimide sensitive fusion (NSF) protein enhancing its ATPase activity and SNARE complex disassembling rate.
LRRK2 phosphorylates pre-synaptic N-ethylmaleimide sensitive fusion (NSF) protein enhancing its ATPase activity and SNARE complex disassembling rate.
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DOI：
10.1186/s13024-015-0066-z
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发表时间：
2016-01-13
影响因子：
15.1
通讯作者：
Greggio E
中科院分区：
文献类型：
--
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
Lrrk 2是一种与帕金森病相关的基因，编码一种大的支架蛋白，具有与囊泡和细胞膜相关过程有关的激酶和GT3活性。在突触前位点，LRRK 2通过与一组突触前蛋白相互作用与突触囊泡相关联。在这里，我们表明，LRRK 2激酶活性影响突触囊泡融合的动力学。因此，我们研究了LRRK 2是否磷酸化胞外/内吞机制的组分。我们先前已经观察到LRRK 2与NSF相互作用，NSF是一种六聚体AAA+ ATP酶，其将ATP水解与SNARE蛋白的分解偶联，使它们在突触胞吐期间进入另一个融合周期。在这里，我们证明了NSF是LRRK 2激酶活性的底物。LRRK 2在D2结构域的ATP结合口袋中的苏氨酸645处磷酸化全长NSF。在功能上，由LRRK 2磷酸化的NSF显示增强的ATP酶活性和增加的SNARE复合物分解速率。用丙氨酸取代苏氨酸645消除了LRRK 2介导的增加的ATP酶活性。鉴于最常见的帕金森病LRRK 2 G2019 S突变显示激酶活性增加，我们的研究结果表明，突变LRRK 2可能通过NSF的异常磷酸化损害突触囊泡动力学。本文的在线版本（doi：10.1186/s13024-015-0066-z）包含补充材料，可供授权用户使用。
Lrrk2, a gene linked to Parkinson’s disease, encodes a large scaffolding protein with kinase and GTPase activities implicated in vesicle and cytoskeletal-related processes. At the presynaptic site, LRRK2 associates with synaptic vesicles through interaction with a panel of presynaptic proteins. Here, we show that LRRK2 kinase activity influences the dynamics of synaptic vesicle fusion. We therefore investigated whether LRRK2 phosphorylates component(s) of the exo/endocytosis machinery. We have previously observed that LRRK2 interacts with NSF, a hexameric AAA+ ATPase that couples ATP hydrolysis to the disassembling of SNARE proteins allowing them to enter another fusion cycle during synaptic exocytosis. Here, we demonstrate that NSF is a substrate of LRRK2 kinase activity. LRRK2 phosphorylates full-length NSF at threonine 645 in the ATP binding pocket of D2 domain. Functionally, NSF phosphorylated by LRRK2 displays enhanced ATPase activity and increased rate of SNARE complex disassembling. Substitution of threonine 645 with alanine abrogates LRRK2-mediated increased ATPase activity. Given that the most common Parkinson’s disease LRRK2 G2019S mutation displays increased kinase activity, our results suggest that mutant LRRK2 may impair synaptic vesicle dynamics via aberrant phosphorylation of NSF. The online version of this article (doi:10.1186/s13024-015-0066-z) contains supplementary material, which is available to authorized users.