Regulation of neurotransmitter release by C2-domain proteins

C2 结构域蛋白对神经递质释放的调节

• 批准号: 9037057
• 负责人: Jose Rizorey
• 金额: $ 44.79万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-25 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9037057
• 关键词: Affect; Area; Binding; Biochemical; Biological Assay; Brain; C2 Domain; Calmodulin; Cell membrane; Characteristics; Communication; Complement; Complex; Crystallization; DAG/PE-Binding Domain; Data; Development; Diglycerides; Disease; Docking; Event; Exhibits; Fluorescence Spectroscopy; Funding; Genetic; Goals; Grant; Health; Knowledge; Laboratories; Ligands; Liposomes; Mediating; Membrane; Membrane Fusion; Modeling; Molecular; Monitor; Monomeric GTP-Binding Proteins; Mutagenesis; NMR Spectroscopy; Neurons; Pharmaceutical Preparations; Phosphatidylinositol 4,5-Diphosphate; Phospholipids; Physiological; Play; Process; Protein Isoforms; Proteins; Regulation; Research; Resolution; S-nitro-N-acetylpenicillamine; SNAP receptor; Structure; Synaptic Transmission; Synaptic Vesicles; Testing; Time; Vesicle; Work; Zinc Fingers; base; controlled release; fascinate; information processing; insight; intermolecular interaction; nervous system disorder; neurotransmitter release; novel strategies; presynaptic; reconstitution; research study; sensor; success; synaptotagmin; synaptotagmin I; syntaxin 1; therapy development; urinary gonadotropin fragment; vesicle-associated membrane protein

DESCRIPTION (provided by applicant): Neurotransmitter release is acutely triggered by Ca2+ and is regulated during presynaptic plasticity processes that underlie some forms of information processing in the brain. Characterization of the mechanisms of release and its regulation is thus critical to understand brain function and will facilitate the development of therapies for neurological disorders with a presynaptic origin. The machinery that controls release contains a core formed by Munc18-1 and SNARE proteins, and specialized proteins that regulate release. Several of these proteins contain multiple C2 domains, which are widespread Ca and phospholipids binding modules but can 2+ also exhibit Ca2+-independent activities. These proteins include: i) Synaptotagmin-1, the Ca2+ sensor that triggers fast release; ii) Munc13-1 and related isoforms, which are essential for release and mediates diverse forms of presynaptic plasticity; and iii) ¿RIMs, which are Rab3 effectors that also have key roles in release and presynaptic plasticity. The C2 domains of all these proteins are highly conserved and are hypothesized in this proposal to regulate neurotransmitter release at multiple levels through their Ca2+-dependent and Ca2+- independent interactions. The ultimate goals of the research proposed in this application are to test this hypothesis and, more generally, to elucidate the molecular mechanisms underlying key forms of regulation of neurotransmitter release. This research forms part of an integrated approach where the structural and reconstitution data performed in the PI's lab are correlated with genetic and physiological experiments performed in the laboratories of close collaborators. We propose three Specific Aims that focus on the following areas: 1. Mechanism of action of Synaptotagmin-1; 2. Structural basis for Munc13-dependent regulation of neurotransmitter release; and 3. Reconstitution of Munc13-dependent regulation of neurotransmitter release. The proposed experiments will build on exciting results obtained during the previous funding period, including a preliminary structure of a Synaptotagmin-1/SNARE complex, key advances in crystallization of large Munc13-1 fragments, the reconstitution of synaptic vesicle fusion with the eight most central components of the release machinery, and preliminary data that reinforce the notion that Munc13s act as master regulators of release through a fascinating network of intramolecular and intermolecular interactions. We expect that this research will provide critical insights into the regulation of release in varied presynaptic plasticity processes, helping to establish fundamental principles on neuronal communication that are vital for brain function.

描述（由申请人提供）：神经递质释放由Ca 2+急性触发，并在突触前可塑性过程中受到调节，该过程是大脑中某些形式的信息处理的基础。因此，释放及其调节机制的表征对于理解脑功能至关重要，并将促进突触前起源的神经系统疾病的治疗方法的发展。控制释放的机制包含由Munc 18 -1和SNARE蛋白形成的核心，以及调节释放的专门蛋白。这些蛋白质中有几种含有多个C2结构域，它们是广泛的Ca和磷脂结合模块，但也可以表现出Ca 2+非依赖性活性。这些蛋白质包括：i）Synaptotagmin-1，触发快速释放的Ca 2+传感器; ii）Munc 13 -1和相关的同种型，它们对释放至关重要，并介导不同形式的突触前可塑性; iii）RIMs，它们是Rab 3效应子，在释放和突触前可塑性中也起关键作用。所有这些蛋白质的C2结构域都是高度保守的，并且在该提议中假设通过其Ca 2+依赖性和Ca 2+非依赖性相互作用在多个水平上调节神经递质释放。本申请中提出的研究的最终目标是测试这一假设，更一般地说，是阐明神经递质释放调节的关键形式的分子机制。这项研究是综合方法的一部分，其中PI实验室中进行的结构和重建数据与密切合作者实验室中进行的遗传和生理实验相关。我们提出了三个具体目标，重点放在以下几个方面：1。突触结合蛋白-1的作用机制; 2. Munc 13依赖性调节神经递质释放的结构基础;和3. Munc 13依赖性调节神经递质释放的重建。拟议的实验将建立在前一个资助期内获得的令人兴奋的结果的基础上，包括Synaptotagmin-1/SNARE复合物的初步结构，大型Munc 13 -1片段结晶的关键进展，突触囊泡融合与释放机制的八个最核心成分的重建，和初步的数据，加强了这一概念，即Munc 13通过一个迷人的分子内和分子间相互作用的网络作为释放的主调节器。我们期望这项研究将为不同突触前可塑性过程中的释放调节提供重要的见解，有助于建立对大脑功能至关重要的神经元通信的基本原则。

项目成果

Subtle Interplay between synaptotagmin and complexin binding to the SNARE complex.

• DOI: 10.1016/j.jmb.2013.07.001
• 发表时间: 2013-09-23
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Xu, Junjie;Brewer, Kyle D.;Perez-Castillejos, Raquel;Rizo, Josep
• 通讯作者: Rizo, Josep

The Janus-faced nature of the C(2)B domain is fundamental for synaptotagmin-1 function.

C（2）B结构域的Janus面对面性质对于SynaptoTagmin-1函数是基础。

• DOI: 10.1038/nsmb.1508
• 发表时间: 2008-11
• 期刊: NATURE STRUCTURAL & MOLECULAR BIOLOGY
• 影响因子: 16.8
• 作者: Xue, Mingshan;Ma, Cong;Craig, Timothy K.;Rosenmund, Christian;Rizo, Josep
• 通讯作者: Rizo, Josep

Munc13 C2B domain is an activity-dependent Ca2+ regulator of synaptic exocytosis.

• DOI: 10.1038/nsmb.1758
• 发表时间: 2010-03
• 期刊: Nature structural & molecular biology
• 影响因子: 16.8

Structure and Ca²⁺-binding properties of the tandem C₂ domains of E-Syt2.

• DOI: 10.1016/j.str.2013.11.011
• 发表时间: 2014-02-04
• 期刊: STRUCTURE
• 影响因子: 5.7
• 作者: Xu, Junjie;Bacaj, Taulant;Zhou, Amy;Tomchick, Diana R.;Suedhof, Thomas C.;Rizo, Josep
• 通讯作者: Rizo, Josep

The C2B domain of synaptotagmin I is a Ca2+-binding module.

• DOI: 10.1021/bi010340c
• 发表时间: 2001-05
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: J. Ubach;Y. Lao;I. Fernández;D. Araç;T. Südhof;J. Rizo