Activity-dependent bulk endocytosis proteome reveals a key presynaptic role for the monomeric GTPase Rab11.
Activity-dependent bulk endocytosis proteome reveals a key presynaptic role for the monomeric GTPase Rab11.
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DOI：
10.1073/pnas.1809189115
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发表时间：
2018-10-23
影响因子：
11.1
通讯作者：
Cousin MA
中科院分区：
文献类型：
--
作者：
Kokotos AC;Peltier J;Davenport EC;Trost M;Cousin MA
通过突触囊泡（SV）再循环维持神经传递对脑功能至关重要。在强烈活动期间，SV的主要再循环模式是活动依赖的大量内吞作用（ADBE），这表明它将在神经传递中发挥关键作用。然而，ADBE的作用仍然是不确定的，由于缺乏识别的分子特异性的这一过程。大量内体蛋白质组（一个关键的ADBE细胞器）的测定显示，它具有独特的分子特征，并确定了Rab11在突触前功能中的作用。这项工作提供了ADBE的分子清单，这对希望在健康和疾病的强烈神经元活动期间调节神经传递的研究人员具有重要价值。活动依赖性大量内吞作用（ADBE）是高频刺激时突触囊泡内吞作用的主要模式，这表明它在神经元强烈活动期间的神经传递中起着关键作用。然而，在阐明ADBE的生理作用的努力受到了阻碍，缺乏识别的分子，这是独特的这种内吞模式。为了解决这一问题，我们对纯化的本体内体进行了蛋白质组学分析，这是ADBE中的关键细胞器。通过两种独立的方法，经典的亚细胞分级分离方法和通过磁性纳米颗粒的分离，富集散装内体。通过两种方案鉴定的蛋白质有77%的重叠，这些分子形成了ADBE核心蛋白质组。生物信息学分析显示，除了预期的突触和运输蛋白外，细胞粘附和细胞骨架和信号分子也有很强的富集。网络分析确定Rab GTP酶作为ADBE蛋白质组内的中心枢纽。随后对这些Rabs的一个子集的研究表明Rab11既促进ADBE又加速网格蛋白介导的内吞作用。这些结果表明，ADBE蛋白质组将为未来的突触前功能的研究提供丰富的资源，并确定Rab11作为突触前功能的调节剂。
The maintenance of neurotransmission by synaptic vesicle (SV) recycling is critical to brain function. The dominant SV recycling mode during intense activity is activity-dependent bulk endocytosis (ADBE), suggesting it will perform a pivotal role in neurotransmission. However, the role of ADBE is still undetermined, due to the absence of identified molecules specific for this process. The determination of the bulk endosome proteome (a key ADBE organelle) revealed that it has a unique molecular signature and identified a role for Rab11 in presynaptic function. This work provides the molecular inventory of ADBE, a resource that will be of significant value to researchers wishing to modulate neurotransmission during intense neuronal activity in both health and disease. Activity-dependent bulk endocytosis (ADBE) is the dominant mode of synaptic vesicle endocytosis during high-frequency stimulation, suggesting it should play key roles in neurotransmission during periods of intense neuronal activity. However, efforts in elucidating the physiological role of ADBE have been hampered by the lack of identified molecules which are unique to this endocytosis mode. To address this, we performed proteomic analysis on purified bulk endosomes, which are a key organelle in ADBE. Bulk endosomes were enriched via two independent approaches, a classical subcellular fractionation method and isolation via magnetic nanoparticles. There was a 77% overlap in proteins identified via the two protocols, and these molecules formed the ADBE core proteome. Bioinformatic analysis revealed a strong enrichment in cell adhesion and cytoskeletal and signaling molecules, in addition to expected synaptic and trafficking proteins. Network analysis identified Rab GTPases as a central hub within the ADBE proteome. Subsequent investigation of a subset of these Rabs revealed that Rab11 both facilitated ADBE and accelerated clathrin-mediated endocytosis. These findings suggest that the ADBE proteome will provide a rich resource for the future study of presynaptic function, and identify Rab11 as a regulator of presynaptic function.
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影响因子：
4.3
作者：
Eden E;Lipson D;Yogev S;Yakhini Z
通讯作者：
Yakhini Z
影响因子：
5.3
作者：
Atluri, PP;Ryan, TA
通讯作者：
Ryan, TA
DOI：
10.1523/jneurosci.4697-12.2013
发表时间：
2013-02-20
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Cheung G;Cousin MA
通讯作者：
Cousin MA
DOI：
10.1523/jneurosci.1976-09.2009
发表时间：
2009-06-17
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Clayton EL;Anggono V;Smillie KJ;Chau N;Robinson PJ;Cousin MA
通讯作者：
Cousin MA
突触时的钙粘蛋白和Catenin：突触发生和突触可塑性中的作用。
影响因子：
15.9
作者：
Arikkath, Jyothi;Reichardt, Louis F.
通讯作者：
Reichardt, Louis F.