Structural Biology of Presynaptic Scaffolds

突触前支架的结构生物学

• 批准号: 7359235
• 负责人: VINZENZ UNGER
• 金额: $ 16.53万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7359235
• 关键词: Address; Amphetamines; Architecture; Binding Proteins; Biology; Brain; Brain Part; Cell physiology; Cellular Membrane; Chromosome Pairing; Cocaine; Complex; Cryoelectron Microscopy; Cues; Cytoplasm; Cytoskeleton; DNA Sequence Rearrangement; Data; Dynamin; Endocytosis; Exposure to; Figs - dietary; Generations; Goals; Guanosine Triphosphate Phosphohydrolases; Image; Ion Channel; Left; Length; Letters; Light; Lipid Bilayers; Lipid Binding; Macromolecular Complexes; Membrane; Molecular; Numbers; Play; Process; Property; Protein Binding; Proteins; Publishing; Recruitment Activity; Recycling; Roentgen Rays; Role; Sampling; Signaling Molecule; Staging; Structure; Surface; Synapses; Synaptic Transmission; Synaptic plasticity; TRIP10 gene; Techniques; Tertiary Protein Structure; Vertebral column; Vesicle; Wiskott-Aldrich Syndrome; Work; addiction; amphiphysin; concept; dimer; drug of abuse; improved; insight; molecular scale; nervous system disorder; particle; presynaptic; receptor; reconstruction; relating to nervous system; scaffold; stem; structural biology; synaptic function

DESCRIPTION (provided by applicant): The macroscopic morphological changes in spine structure, and number of synapses that accompany exposure to drugs of abuse such as cocaine and amphetamines indicate that synaptic plasticity plays an important role in the re-wiring of an addicted brain. Descending to a molecular level, cues for changes in the circuitry are issued through changes in synaptic activity, which critically depends on the cellular process of endocytosis because of its direct involvement in receptor clearance from the surface and neurotramsitter vesicle/transporter recycling. The goal of this exploratory study is to enable mechanistic understanding of fundamental synaptic processes, such as endocytosis, by opening the dynamic interface between cellular membranes and the cytoplasm to structural exploration. As a starting point, our work will focus on BAR- and F-BAR-domain proteins whose ability to induce, and to stabilize membrane curvature places them at a strategic intersection between membrane remodeling, vesicle fission, and cytoskeletal re-arrangements. Using purified BAR-/F-BAR-domains and their corresponding full-length proteins (endophilin, amphiphysin, syndapin 1, FBP17 and CIP4), we seek to obtain samples for structure determination by electron cryomicroscopy, in which these proteins are bound to lipid bilayers, either alone or in complex with downstream interaction partners like the fission GTPase dynamin, the cytoskeletal regulators Cdc42 and neural Wiskott-Aldrich-Syndrome Protein (N-WASP). Exposure to drugs of abuse such as cocaine and amphetamines results in lasting morphological changes in some parts of the brain, indicating that synaptic plasticity plays an important role in the biology of addiction. At the molecular level, cues for remodeling of synapses stem from changes in synaptic activities, which at the molecular level critically depend on the cellular process of endocytosis. To advance our basic understanding of the complex processes underlying the biology of addiction, this study ultimately aims at revealing, at a molecular scale, the mechanisms through which protein complexes that are critically important for endocytotic processes contribute to synaptic plasticity.

描述（由申请人提供）：脊柱结构的宏观形态学变化以及伴随暴露于滥用药物（如可卡因和安非他明）的突触数量表明突触可塑性在成瘾大脑的重新布线中起重要作用。下降到分子水平，在电路中的变化的线索是通过突触活动的变化，这在很大程度上取决于细胞内吞作用的过程，因为它直接参与受体清除表面和神经递质囊泡/转运蛋白再循环。这项探索性研究的目标是通过打开细胞膜和细胞质之间的动态界面进行结构探索，从而实现对基本突触过程（如内吞作用）的机械理解。作为一个起点，我们的工作将集中在BAR和F-BAR结构域蛋白的诱导能力，并稳定膜曲率的地方，他们在膜重塑，囊泡分裂和细胞骨架重排之间的战略交叉点。使用纯化的BAR-/F-BAR-结构域及其相应的全长蛋白（内亲蛋白、双端外膜蛋白、syndapin 1、FBP 17和CIP 4），我们试图获得用于通过电子冷冻显微镜进行结构测定的样品，其中这些蛋白质单独或与下游相互作用配偶体（如裂变GT3发动蛋白）复合地结合至脂质双层，细胞骨架调节因子Cdc 42和神经Wiskott-Aldrich综合征蛋白（N-WASP）。接触可卡因和安非他明等滥用药物会导致大脑某些部位发生持久的形态学变化，这表明突触可塑性在成瘾生物学中起着重要作用。在分子水平上，突触重塑的线索源于突触活动的变化，而突触活动在分子水平上严重依赖于细胞内吞过程。为了推进我们对成瘾生物学基础的复杂过程的基本理解，本研究的最终目的是在分子水平上揭示对内吞过程至关重要的蛋白质复合物有助于突触可塑性的机制。