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变化，以及伴随滥用药物（例如可卡因和苯丙胺）的突触数量，表明突触可塑性在重新培养上瘾的大脑的重新培训中起着重要作用。下降到分子水平，通过突触活动的变化发出了电路变化的线索，由于其直接参与了表面和神经剂量囊泡/转运蛋白回收的受体清除率，因此在关键上取决于内吞作用的细胞过程。这项探索性研究的目的是通过将细胞膜和细胞质之间的动态界面开放到结构探索中，以实现对基本突触过程（例如内吞作用）的机械理解。作为起点，我们的工作将集中在棒和F型杆域蛋白上，其诱导能力并稳定膜曲率将它们置于膜重塑，囊泡裂变和细胞骨架重组之间的战略相交。我们使用纯化的棒/F-BAR-层及其相应的全长蛋白（内po蛋白，两亲性，联合辛1，FBP17和CIP4），我们试图通过电子酸球镜检查来确定结构的样品，这些蛋白与这些蛋白具有与脂肪双层相比，在其中涉及这些蛋白质的依次，或者与下层相互作用，而彼此中的相互作用是彼得的互动，该蛋白质与下层相互作用相比，细胞骨架调节剂CDC42和神经Wiskott-Aldrich-Syndrome蛋白（N-WASP）。可卡因和苯丙胺等滥用药物的暴露会导致大脑某些部位的形态变化，这表明突触可塑性在成瘾的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，突触活动在分子水平上严重取决于内吞作用的细胞过程。为了促进我们对成瘾生物学生物学基础的复杂过程的基本理解，这项研究最终旨在以分子规模揭示对内吞过程至关重要的蛋白质复合物的机制，从而有助于突触可塑性。