FUNCTIONS OF NEURONAL PENTRAXINS AND TCBP49

神经元五聚蛋白和 TCBP49 的功能

• 批准号: 6540182
• 负责人: MARK S PERIN
• 金额: $ 25.9万
• 依托单位: CLEVELAND CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6540182
• 关键词: CHO cells; PC12 cells; binding proteins; calcium binding protein; clathrin; endocytosis; glutamate receptor; laboratory mouse; laboratory rat; nerve /myelin protein; neurochemistry; neurons; neurotoxins; protein biosynthesis; protein localization; protein protein interaction; protein structure function; synapses; synaptogenesis

The aim of this application is the elucidation of the functional role of four novel binding proteins for a presynaptic-acting snake venom toxin, taipoxin, that blocks synaptic vesicle recycling. Three of these proteins (neuronal pentraxin 1, NP1; neuronal pentraxin 2, NP2; and neuronal pentraxin receptor, NPR) have homology to acute phase proteins, C-reactive protein and serum amyloid P protein that recognize bacteria and cell debris. We propose that these neuronal pentraxins represent a novel neuronal uptake pathway that may be associated with synaptic formation and remodeling and potentially AMPA receptor clustering. The fourth protein is a novel reticular calcium- binding protein, taipoxin-associated calcium binding protein 49 (TCBP49). To test the function of these proteins and to test our hypothesis that these proteins form a pathway for synaptic uptake, we will undertake the following experiments. First, we will test if all neuronal pentraxins are localized to synapses and whether their message expression follows synaptogenesis. Second, we will test for alterations of synapses, synaptic physiology and taipoxin action in mice that lack neuronal pentraxins. Third, we will test the role of TCBP49, NP1, NP2 and NPR in taipoxin blockade of endocytosis in CHO and PC12 cells transfected to express these proteins or express antisense RNA. Fourth, we will expand our chromatographies to investigate proteins that interact with glycosylated, homo- and heteropentameric NP1, NP2 and NPR to test the connection of these proteins to the endocytic AP-2 clathrin adaptor complex and to AMPA receptors.

该应用的目的是阐明四种新型结合蛋白在突触前作用蛇毒毒素Taipoxin的功能作用，从而阻断突触囊泡回收。 这些蛋白质中的三个（神经元五肽1，NP1；神经元五肽2，NP2；和神经元五肽受体，NPR）与急性相蛋白，C-反应蛋白和血清蛋白P蛋白具有识别细菌和细胞Debris的C蛋白具有同源性。我们建议这些神经元五肽代表一种新型的神经元摄取途径，可能与突触形成和重塑以及潜在的AMPA受体聚类有关。 第四蛋白是一种新型的网状钙结合蛋白，taipoxin相关的钙结合蛋白49（TCBP49）。 为了测试这些蛋白质的功能，并测试了我们的假设，即这些蛋白质构成了突触摄取的途径，我们将进行以下实验。 首先，我们将测试是否将所有神经元五生型都定位于突触以及它们的信息表达是否遵循突触发生。其次，我们将测试缺乏神经元五肽的小鼠突触的改变，突触生理和taipoxin作用。 第三，我们将测试TCBP49，NP1，NP2和NPR在CHO和PC12细胞中转染以表达这些蛋白或表达反义RNA的CHO和PC12细胞中taipoxin封闭中的作用。第四，我们将扩展色谱法以研究与糖基化，同型和异源性NP1，NP2和NPR相互作用的蛋白质，以测试这些蛋白质与内吞AP-2网格蛋白衔接子复合物和AMPA受体的连接。