STRUCTURE/FUNCTION OF NERVE GROWTH FACTOR/NEUROTROPHINS

描述(申请人摘要)：神经营养因子在 发展和维持多种类型的神经元。神经家族 生长因子(NGF)相关神经营养因子(BDNF、NT-3、NT-4/5)与 低亲和力神经营养因子受体(P75NTR)与trk家族 原癌基因受体酪氨酸激酶(Trk)在体内建立信号转导 反应性外周神经元(感觉、交感)和中枢神经元 (胆碱能、多巴胺能)。在此测试的总体假设 建议认为NGF与其受体之间的最初相互作用包括 导致离散的构象变化和受体二聚化 细胞内信号通路可以被适当的单独激活 神经生长因子突变体。这种作用可能导致新的和高度特异的治疗方法 治疗神经性疾病的药物。此应用程序的特定目标是 解决这一假设的方法是：开发能够独立传递信号的NGF突变体 用于分化、生长停滞、细胞凋亡和生存；确定 区分这些突变体的信号通路；并测量 NGF及其突变体与神经生长因子相互作用时的构象变化 TrkA-red和四种神经营养因子与p75NTR-red利用不同的 荧光技术。一般来说，这些项目使用的是纯化的蛋白质 具有物理生化技术的组件和用于信号传递的PC12细胞 分析。所有的NGF突变体都被纯化并鉴定了其结构 正直。分子生物学、免疫化学、光谱和化学 将利用各种方法。预计在这里的结果将有助于为 为有意义和合理的治疗试剂设计奠定基础 来源于选择性刺激信号通路的神经营养因子突变体 并最终可能导致阿尔茨海默病的新疗法， 帕金森病、儿童神经母细胞瘤及相关神经病学 精神错乱。例如，NGF现在已知可以防止中枢神经系统神经元萎缩和 刺激脊髓感觉神经元的再生--我们的 反应选择性NGF突变体最终可能会更有效 治疗这种神经性疾病。

DESCRIPTION (Applicant's abstract): Neurotrophic factors play a major role in development and maintenance of many types of neurons. The family of nerve growth factor (NGF) related neurotrophins (BDNF, NT-3, NT-4/5) interact with a low affinity neurotrophin receptor (p75NTR) and with the family of trk proto-oncogene receptor tyrosine kinases (Trk) to establish signaling within responsive peripheral neurons (sensory, sympathetic) and central neurons (cholinergic, dopaminergic). The overall hypothesis being tested in this proposal is that the initial interaction between NGF and its receptors involves a conformational change and receptor dimerization that leads to discrete intracellula signaling pathways that can be separately activated by appropriate NGF mutants. Such effects could lead to novel and highly specific therapeutic agents in neurological disorders. The Specific Aims of this application that address this hypothesis are: to develop NGF mutants that independently signal for differentiation, growth arrest, apoptosis and survival; to determine the signaling pathways that distinguish these mutants; and to measure conformational changes upon interaction of NGF and the mutants with the TrkA-RED and of the four neurotrophins with the p75NTR-RED utilizing various fluorescence techniques. In general, these projects utilize purified protein components with physical biochemical techniques and PC12 cells for signaling analysis. All NGF mutants are purified and characterized for structural integrity. Molecular biological, immunochemical, spectroscopic, and chemical approaches will be utilized. The results expected here will help lay the foundation for the meaningful and rational design of therapeutic reagents derived from neurotrophin mutants that selectively stimulate signaling pathways and could eventually lead to novel treatments of Alzheimer's Disease, Parkinson's Disease, childhood neuroblastomas, and related neurological disorders. For example, NGF is now known to prevent atrophy 0 CNS neurons and to stimulate regeneration of sensory neurons in the spinal cord - our response-selective NGF mutants may be even more effective in ultimately treating such neurological disorders.