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通过涉及变量N终端区域的自抑制相互作用控制Munc13-1活动

Junjie Xu^1,2,3, Victoria Esser^1,2,3, Katarzyna Gołębiowska-Mendroch^1,2,3,4

¹Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.

bioRxiv : the preprint server for biology

|February 8, 2024

概括

神经递质释放中的Munc13-1蛋白活性由其N端域调节. 阿尔法-RIM和卡尔莫杜林通过与Munc13-1结合来缓解抑制,从而使大脑正常运作.

科学领域:

神经科学是一个神经科学.
分子生物学分子生物学
生物化学生物化学

背景情况:

预突触可塑性对于大脑信息处理至关重要,它依赖于受调节的神经递质释放.
Munc13蛋白,特别是Munc13-1,是通过其C端 MUN 域和可变的N端区域释放神经递质的关键调节者.
Munc13-1的N终端区域包含一个卡尔莫杜林结合 (CaMb) 域和一个C2A域,它们与αRIM蛋白相互作用以调节可塑性.

研究的目的:

研究Munc13-1的N和C终端区域的综合功能.
描述Munc13-1活动的调节机制,涉及其N端域,αRIM和calmodulin.
为了阐明这些相互作用如何影响SNARE复合体组合和神经递质释放.

主要方法:

纯化一个全长的Munc13-1片段,包括MUN,C2A和CaMb域.
脂质体融合试验测量蛋白质在促进膜融合中的活性.
核磁共振 (NMR) 光谱法用于确定域相互作用.

主要成果:

净化后的Munc13-1片段与单独的C端区域相比,其活性降低.
通过添加RIM2α指 (ZF) 域和calmodulin,Munc13-1片段的活性显著增强.
核磁共振数据显示,C2A和CaMb域与MUN域结合,这些抑制相互作用由RIM2αZF和calmodulin分别缓解.

结论:

Munc13-1的活性通过涉及其C2A和CaMb域与MUN域结合的分子内抑制来调节.
αRIM蛋白和calmodulin通过破坏这些抑制相互作用,起到关键的积极调节者的作用.
这为Munc13-1如何集成信号来控制SNARE复合体组合和神经递质释放在突触可塑性期间提供了一个分子模型.

关键词:

神经递质释放神经递质的释放内部分子相互作用.突触前可塑性突触前可塑性蛋白质NMR是NMR的一个蛋白质.突触囊泡融合是指突触囊泡的融合

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