LRRK2 mediated Rab8a phosphorylation promotes lipid storage.
LRRK2 mediated Rab8a phosphorylation promotes lipid storage.
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LRRK2 介导的 Rab8a 磷酸化促进脂质储存
DOI：
10.1186/s12944-018-0684-x
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发表时间：
2018-02-27
影响因子：
4.5
通讯作者：
Zhou L
中科院分区：
文献类型：
--
作者：
Yu M;Arshad M;Wang W;Zhao D;Xu L;Zhou L
富含亮氨酸重复激酶2（LRRK 2）基因的几个突变与帕金森病（PD）的发病机制有关，帕金森病是一种神经退行性疾病，其特征是静息性震颤和僵硬，导致姿势不稳定。研究表明，导致LRRK2蛋白激酶活性增加的突变与PD发病机制显著相关。最近的研究表明，一些Rab GTP酶，特别是Rab8，作为LRRK2的底物，并在相互作用时在其开关II结构域中进行磷酸化。本研究旨在探讨Rab8及其突变体的磷酸化对脂代谢和脂滴生长的影响。Rab8a的磷酸化状态通过荧光标记凝胶检测。产生点突变体构建体以研究Rab8a的功能。用指定的质粒转染3T3L1细胞，并用Bodipy染色脂滴。进行荧光显微镜实验以检查脂滴的大小。通过免疫沉淀和western blot检测Rab8a与Optineurin的相互作用。我们的分析表明Rab8a被具有高激酶活性的突变LRRK2磷酸化。Rab8a在T72氨基酸残基上的磷酸化促进了大脂滴的形成。与野生型Rab8a相比，突变体T72D具有更高的促进大脂滴形成的活性，脂滴的平均直径从2.10 μ m增加到2.46 μ m。此外，Rab8a的磷酸化减弱了与其效应子视神经磷酸酶的相互作用。Y1699C突变的LRRK2能够磷酸化Rab8a，Rab8a在位点72的磷酸化在脂滴的融合和扩大中起重要作用。总之，我们的研究表明，增强脂质储存能力和PD发病机制之间的间接关系。
Several mutations in leucine rich repeat kinase 2 (LRRK2) gene have been associated with pathogenesis of Parkinson’s disease (PD), a neurodegenerative disorder marked by resting tremors, and rigidity, leading to Postural instability. It has been revealed that mutations that lead to an increase of kinase activity of LRRK2 protein are significantly associated with PD pathogenesis. Recent studies have shown that some Rab GTPases, especially Rab8, serve as substrates of LRRK2 and undergo phosphorylation in its switch II domain upon interaction. Current study was performed in order to find out the effects of the phosphorylation of Rab8 and its mutants on lipid metabolism and lipid droplets growth. The phosphorylation status of Rab8a was checked by phos-tag gel. Point mutant construct were generated to investigate the function of Rab8a. 3T3L1 cells were transfected with indicated plasmids and the lipid droplets were stained with Bodipy. Fluorescent microscopy experiments were performed to examine the sizes of lipid droplets. The interactions between Rab8a and Optineurin were determined by immunoprecipitation and western blot. Our assays demonstrated that Rab8a was phosphorylated by mutated LRRK2 that exhibits high kinase activity. Phosphorylation of Rab8a on amino acid residue T72 promoted the formation of large lipid droplets. T72D mutant of Rab8a had higher activity to promote the formation of large lipid droplets compared with wild type Rab8a, with increase in average diameter of lipid droplets from 2.10 μm to 2.46 μm. Moreover, phosphorylation of Rab8a weakened the interaction with its effector Optineurin. Y1699C mutated LRRK2 was able to phosphorylate Rab8a and phosphorylation of Rab8a on site 72 plays important role in the fusion and enlargement of lipid droplets. Taken together, our study suggests an indirect relationship between enhanced lipid storage capacity and PD pathogenesis.
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影响因子：
3.7
作者：
Park B;Ying H;Shen X;Park JS;Qiu Y;Shyam R;Yue BY
通讯作者：
Yue BY
DOI：
10.15252/embj.201591593
发表时间：
2015-11-12
期刊：
The EMBO journal
影响因子：
--
作者：
Lai YC;Kondapalli C;Lehneck R;Procter JB;Dill BD;Woodroof HI;Gourlay R;Peggie M;Macartney TJ;Corti O;Corvol JC;Campbell DG;Itzen A;Trost M;Muqit MM
通讯作者：
Muqit MM
大鼠富含亮氨酸重复激酶 2 (LRRK2) 的缺失会导致外周器官逐渐出现异常表型。
影响因子：
3.7
作者：
Baptista MA;Dave KD;Frasier MA;Sherer TB;Greeley M;Beck MJ;Varsho JS;Parker GA;Moore C;Churchill MJ;Meshul CK;Fiske BK
通讯作者：
Fiske BK
影响因子：
6.1
作者：
Greggio, Elisa;Jain, Shushant;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
影响因子：
3.5
作者：
Gloeckner, CJ;Kinkl, N;Ueffing, M
通讯作者：
Ueffing, M