STRUCTURE/FUNCTION OF NERVE GROWTH FACTOR/NEUROTROPHINS

描述（申请人摘要）：神经营养因子在 许多类型的神经元的发育和维持。神经家族 生长因子（NGF）相关的神经营养因子（BDNF，NT-3，NT-4/5）与一种 低亲和力神经营养因子受体（p75 NTR）和trk家族 原癌基因受体酪氨酸激酶（Trk），以建立信号转导内 反应性外周神经元（感觉神经元、交感神经元）和中枢神经元 （胆碱能，多巴胺能）。在这个实验中， 建议是，最初的相互作用之间的神经生长因子和它的受体涉及 构象变化和受体二聚化，导致离散的 细胞内信号通路，可以分别激活适当的 NGF突变体。这种效应可能会导致新的和高度特异性的治疗 神经系统疾病的药物。本申请的具体目的是 解决这一假设是：开发独立信号的NGF突变体， 用于分化、生长停滞、凋亡和存活;以确定 区分这些突变体的信号通路;并测量 构象变化后的相互作用的神经生长因子和突变体与 TrkA-RED和四种神经营养因子中的p75 NTR-RED利用各种 荧光技术一般来说，这些项目利用纯化的蛋白质 用物理生化技术和PC 12细胞进行信号转导 分析.所有的NGF突变体都是纯化的，并进行了结构表征。 完整分子生物学、免疫化学、光谱学和化学 方法将被使用。这里预期的结果将有助于奠定 治疗试剂有意义和合理设计的基础 源自选择性刺激信号通路的神经营养蛋白突变体 并可能最终导致阿尔茨海默病的新疗法， 帕金森病、儿童神经母细胞瘤和相关神经系统疾病 紊乱例如，现在已知NGF可防止CNS神经元萎缩， 刺激脊髓感觉神经元的再生-我们的 反应选择性NGF突变体可能在最终 治疗这种神经系统疾病。

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.