Small-Molecule Probes for Conditional Modulation of the Synapse Proteome

用于突触蛋白质组条件调节的小分子探针

• 批准号: 7229891
• 负责人: STEPHEN J HAGGARTY
• 金额: $ 26.15万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7229891
• 关键词: Acetylation; Acute; Affinity; Affinity Chromatography; Antibodies; Area; Bacteria; Behavior; Binding; Biochemical Process; Bioinformatics; Biological Assay; Biological Factors; Biology; Brain; Brain Chemistry; Cell Line; Cellular biology; Chemicals; Chemistry; Chimeric Proteins; Chromosome Pairing; Cloning; Collection; Communities; Complementary DNA; Complex; DNA; Databases; Dendrites; Descriptor; Development; Disease; Elements; Fluorescence Microscopy; Fluorescent Dyes; Future; Genes; Genetic Recombination; Genome; Genomics; Goals; Hippocampus (Brain); Image; Immune Sera; In Vitro; Individual; Label; Learning; Length; Long-Term Effects; Mammalian Cell; Measures; Mediating; Memory; Mental disorders; Methodology; Methods; Mining; Modeling; Molecular; Molecular Probes; Monitor; Mus; Nature; Nervous System Physiology; Neuraxis; Neurobiology; Neurons; Neurosciences; Neurotransmitter Receptor; Neurotransmitters; Numbers; Open Reading Frames; Organic Synthesis; Organism; Pathway interactions; Peptides; Phosphorylation; Play; Process; Protein Chemistry; Proteins; Proteome; Proteomics; Recombinants; Reporter; Research; Research Personnel; Resources; Role; Screening procedure; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Solubility; Source; Standards of Weights and Measures; Structure; Surface Plasmon Resonance; Synapses; Synaptic Vesicles; Synaptic plasticity; Synaptosomes; System; Technology; Work; base; chemical genetics; chemical synthesis; computerized tools; expression cloning; immunocytochemistry; improved; insight; nervous system disorder; neurophysiology; neuropsychiatry; optical imaging; programs; protein folding; protein function; small molecule; small molecule libraries; synaptic function; synaptogenesis; tool

DESCRIPTION (provided by applicant): Neuroscience, like many fields, is limited by the technology available to dissect complex cellular and biochemical processes. Further elucidation of neurophysiology requires methodology for the conditional perturbation of individual components of the underlying molecular and cellular networks. This proposal is directed toward the development of small-molecular probes that will allow for the conditional modulation of multifunction, pre- and post-synaptic proteins, and the signal transduction pathways regulated by neurotransmitter. To accomplish this goal we will: 1) assemble, express, and purify a collection of synaptic proteins after using computational tools to identify key sequence elements and functional domains; 2) identify small-molecule probes that selectively bind to each protein by performing over 3 million high-throughput binding assays using small-molecule microarrays composed of chemicals derived diversity-oriented organic synthesis and known bioactives; 3) validate the observed in vitro binding interactions using surface plasmon resonance analysis; and finally, 4) to characterize the phenotypic effects of the small-molecule probes using neuronal cultures, fluorescence microscopy and genetically-encoded fluorescent reporters to measure synapse assembly, synaptic vesicle dynamics, and synaptic signaling. Developing small-molecule probes, and improving the technologies for their discovery, will expand the molecular toolkit available for dissecting complex neuronal processes, including those implicated in neurological disorders and psychiatric disease. Small-molecule probes provide the opportunity to dissect the complex functions of proteins and molecular networks regulating neurophysiology. Besides increasing our understanding of the cell biology of the synapse, these findings will ultimately enable a better understanding of the relationship between brain chemistry and behavior, as well as to better understand the causes of neuropsychiatric and neurological diseases. In the future chemical biology in general, and chemical genetics in particular, will play an important role in catalyzing new discoveries and potentially insight into function of the nervous system.

描述（由申请人提供）：神经科学与许多领域一样，受到可用于剖析复杂细胞和生化过程的技术的限制。 神经生理学的进一步阐明需要对基础分子和细胞网络的各个组成部分进行条件扰动的方法。 该提案旨在开发小分子探针，以实现多功能、突触前和突触后蛋白质的条件调节，以及由神经递质调节的信号转导途径。 为了实现这一目标，我们将：1）在使用计算工具识别关键序列元件和功能域后组装、表达和纯化突触蛋白集合； 2) 使用小分子微阵列进行超过 300 万次高通量结合测定，识别选择性结合每种蛋白质的小分子探针，所述小分子微阵列由化学衍生的面向多样性的有机合成和已知的生物活性物质组成； 3) 使用表面等离子体共振分析验证观察到的体外结合相互作用；最后，4) 使用神经元培养物、荧光显微镜和基因编码荧光报告基因来表征小分子探针的表型效应，以测量突触组装、突触小泡动力学和突触信号传导。 开发小分子探针并改进其发现技术，将扩大可用于解剖复杂神经元过程的分子工具包，包括那些与神经系统疾病和精神疾病有关的神经元过程。 小分子探针提供了剖析调节神经生理学的蛋白质和分子网络的复杂功能的机会。 除了增加我们对突触细胞生物学的理解之外，这些发现最终将使我们能够更好地理解大脑化学和行为之间的关系，以及更好地理解神经精神和神经系统疾病的原因。 未来，化学生物学，特别是化学遗传学，将在催化新发现和深入了解神经系统功能方面发挥重要作用。