MOLECULAR MECHANISMS UNDERLYING GAP-43 FUNCTION

GAP-43 功能的分子机制

• 批准号: 2857470
• 负责人: RACHAEL Lee NEVE
• 金额: $ 24.24万
• 依托单位: MCLEAN HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2857470
• 关键词: PC12 cells; calmodulin; confocal scanning microscopy; endocytosis; enzyme activity; guanine nucleotide binding protein; intermolecular interaction; laboratory mouse; membrane fusion; membrane proteins; neural growth associated protein; neurogenesis; neurotransmitter transport; protein kinase C; protein structure function

DESCRIPTION: A large body of evidence implicates the neuronal growth associated protein GAP-43 in the modulation of neuronal plasticity during both development and learning. However, the molecular mechanisms by which GAP-43 controls these functions are not known. The present proposal uses analyses of functional domains of GAP-43, coupled with studies of its interaction with rabaptin-5, a protein that mediates endocytic membrane fusion, to delineate the molecular events underlying GAP-43 participation in neurite outgrowth ad neurotransmitter release. First, the hypothesis that specific domains of GAP-43 can be linked to specific signaling pathways and to its roles in the distinct processes of neurotransmitter release and neurite outgrowth will be tested. GAP-43 mutants that disrupt functional domains of the protein will be expressed in PC12 cells and neurons lacking GAP-43. The interactions of these mutants with calmodulin, protein kinase C, and the GTP-binding protein G0 will be evaluated, following which the effects of these mutations on neurotransmitter release and process outgrowth will be quantified. The second hypothesis to be tested is that the known interactions of GAP-43 with calmodulin and protein kinase C are tied to its interactions with rabaptin-5 in the presynaptic terminal. Domain mapping, co-localization studies, and quantitative functional assays will clarify the functional role of the binding of GAP-43 to rabaptin-5, and demonstrate whether endocytic membrane fusion events involving these molecules comprise a molecular link between neurotransmitter release and neurite outgrowth. The data obtained from the proposed studies will begin to provide unifying molecular mechanisms for the diverse functions in which GAP-43 is known to participate.

描述：大量证据暗示了神经元的生长 相关蛋白质GAP-43在调节神经元可塑性中的相关蛋白质GAP-43 发展和学习。 但是，分子机制 GAP-43控制这些功能尚不清楚。 目前的建议使用 GAP-43功能结构域的分析，再加上其研究域的研究 与Rabaptin-5的相互作用，Rabaptin-5，一种介导内吞膜的蛋白质 融合，描绘GAP-43参与的分子事件 神经突生长AD神经递质释放。 首先，假设 GAP-43的特定域可以链接到特定的信号通路和 它在神经递质释放的不同过程中的作用 神经突生长将进行测试。 GAP-43突变体，破坏功能 蛋白质的结构域将在PC12细胞和缺乏神经元中表达 GAP-43。 这些突变体与钙调蛋白，蛋白激酶的相互作用 C和GTP结合蛋白G0将进行评估，然后将其 这些突变对神经递质释放和过程产物的影响 将被量化。 要检验的第二个假设是已知 GAP-43与钙调蛋白和蛋白激酶C的相互作用与其相关 与突触前末端中的Rabaptin-5相互作用。 域映射， 共定位研究和定量功能测定将阐明 GAP-43与Rabaptin-5结合的功能作用，并证明 涉及这些分子的内吞膜融合事件是否包括 神经递质释放与神经突生长之间的分子联系。 从拟议的研究中获得的数据将开始提供统一 已知GAP-43的各种功能的分子机制 参加。