Structural Characterization of Endophilin-2

Endophilin-2 的结构表征

• 批准号: 7284691
• 负责人: Evelyn J. Swain
• 金额: $ 4.45万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7284691
• 关键词: SDS polyacrylamide gel electrophoresis; X ray crystallography; crystallization; endocytosis; postdoctoral investigator; protein binding; protein purification; protein quantitation /detection; protein structure; proteolysis; recombinant proteins

DESCRIPTION (provided by applicant): The long-term goal of this research is to determine the crystal structures of various constructs of endophilin-A2, which will augment existing in vivo and molecular biology studies of the mechanisms involved in the complex and highly regulated cellular process of endocytosis. Endophilins are an SH3 domain containing family of proteins that function in processes such as clathrin-mediated endocytosis, protein trafficking, cell architecture maintenance, cell proliferation and differentiation, and in the immune response. There are no available structural data for these proteins. In this proposal we will purify recombinant rat endophilin-A2 from E. coli and initiate crystallization screens. To facilitate protein crystallization we will identify stable domains by limited proteolysis. Finally, crystal growth will be optimized to generate welldiffracting crystals to allow for structural determination studies. Structural information will lead to insights into how endophilin-A2 functions, both alone and with its various protein binding partners. This will increase our understanding of basic endocytic processes, and how alterations in these processes can result in disease states.

描述（由申请人提供）： 这项研究的长期目标是确定内菲尔蛋白A2的各种构建体的晶体结构，该结构将增强体内现有的现有和分子生物学研究，以了解与内吞作用的复杂且高度调节的细胞过程相关的机制。内芬蛋白是一个含有蛋白质家族的SH3结构域，在诸如网格蛋白介导的内吞作用，蛋白质运输，细胞结构维持，细胞增殖和分化以及免疫反应等过程中起作用。这些蛋白质没有可用的结构数据。在此提案中，我们将从大肠杆菌中纯化重组大鼠内po蛋白-A2，并开始结晶筛。为了促进蛋白质结晶，我们将通过有限的蛋白水解鉴定稳定的结构域。最后，将优化晶体生长以产生良好的晶体，以进行结构测定研究。结构信息将导致有关单独及其各种蛋白质结合伴侣的内pophilin-A2如何功能的见解。这将增加我们对基本内吞过程的理解，以及这些过程中的变化如何导致疾病状态。