NERVE GROWTH FACTOR RECEPTOR--A MOLECULAR ANALYSIS

神经生长因子受体——分子分析

• 批准号: 3079847
• 负责人: BARBARA L HEMPSTEAD
• 金额: $ 6.39万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1991-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3079847
• 关键词: affinity chromatography; cell differentiation; cell growth regulation; genetic manipulation; growth factor receptors; growth inhibitors; hormone receptor; ligands; membrane proteins; molecular cloning; molecular oncology; molecular pathology; neoplastic cell culture for noncancer research; neural crest; neuroblastoma; neurotrophic factors; nucleic acid probes; polyethylenes; transfection

Nerve growth factor (NGF) is an important growth regulatory polypeptide which promotes the survival and differentiation of immature neuroblasts in the developing nervous system. Unlike most growth hormones which stimulate cell replication, NGF inhibits cell proliferation and induces the maturation of cells bearing specific NGF receptors. Thus, the NGF-NGF receptor interaction serves as an important model system to use in investigating the normal mechanisms which regulate cell growth and differentiation. Furthermore, since the effects of NGF extend even to neoplastic cells bearing functional receptors, NGF is of potential clinical significance as a growth inhibitor of neoplasias of neural crest origin. Although NGF was the first growth factor to be identified, its mechanism of action is not well understood, except that its actions are initiated by binding to a high affinity cell surface receptor. This proposal is directed towards defining the role of the NGF receptor in initiating cellular responses to NGF, using the recently cloned gene that encodes the human NGF receptor. A combined molecular biological and biochemical approach will be used to determine the structure of the functionally active high affinity receptor. Specifically we plan to: (a) transfect the human NGF receptor clone into cells derived from the neural crest to determine the cellular characteristics which are responsible for responsiveness to NGF; (b) define the molecular basis of the high and low affinity forms of the receptor by membrane fusion experiments using PEG-mediated fusion; (c) define the domains of the receptor responsible for high affinity ligand binding, internalization and interactions with cell membrane proteins; (d) biochemically characterize the high affinity receptor complex using affinity chromatography and (e) use gene transfer to identify other gene products in the cell membrane which are required for functionally active high affinity receptor.

神经生长因子（NGF）是一种重要的生长调节因子 促进细胞存活和分化的多肽 发育中的神经系统中未成熟的神经母细胞。 不像 大多数刺激细胞复制的生长激素，NGF 抑制细胞增殖并诱导细胞成熟 具有特定的 NGF 受体。 因此，NGF-NGF受体 交互作为一个重要的模型系统用于 研究调节细胞生长的正常机制 和差异化。 此外，由于 NGF 的作用 甚至延伸至带有功能性受体 NGF 的肿瘤细胞 作为生长抑制剂具有潜在的临床意义 神经嵴起源的肿瘤。 尽管 NGF 是第一个被发现的生长因子，但它 其作用机制尚不十分清楚，除了其 通过与高亲和力的细胞表面结合来启动作用 受体。 该提案旨在明确以下机构的作用： NGF 受体启动细胞对 NGF 的反应，使用 最近克隆的编码人类 NGF 受体的基因。 分子生物学和生化相结合的方法将 用于确定功能活性高的结构 亲和力受体。 具体来说，我们计划：（a）转染 将人 NGF 受体克隆到神经细胞衍生的细胞中 波峰以确定细胞特征 负责对 NGF 的反应； (b) 定义分子 受体的高亲和力和低亲和力形式的基础 使用 PEG 介导的融合进行膜融合实验； (三) 定义负责高亲和力的受体的结构域 配体结合、内化以及与细胞的相互作用 膜蛋白； (d) 生物化学特征 使用亲和层析的亲和受体复合物和(e) 使用基因转移来鉴定细胞中的其他基因产物 功能活性高亲和力所需的膜 受体。