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STRUCTURAL ASPECTS OF APP FUNCTION AND PATHOLOGY

应用程序功能和病理学的结构方面

基本信息

批准号：
2732539
负责人：
STEPHEN P ANDERSON
金额：
$ 23.91万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-09-30 至 2000-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2732539
关键词：
amyloid proteins binding proteins high performance liquid chromatography intermolecular interaction plasminogen plasminogen activator protein structure function serine proteinases site directed mutagenesis thrombin

项目摘要

DESCRIPTION (Adapted from the applicant's abstract) The long-term objective of this application is determination of the three dimensional structure of APP by analysis of the structures of its individual functional domains. The cysteine-rich N-terminal domain has been expressed at high levels and purified to homogeneity. Optimization of expression constructs will be required for production of material suitable for X-ray crystallography and NMR. In order to better understand the physiological behavior of APP -- both in normal tissue and in the context of Alzheimer's disease -- attempts are being made to identify APP binding proteins in the body. The concentration is on isolating potential proteases that interact with the APP Kunitz inhibitor (APP-KI) domain. Successful cloning of one such candidate protease utilizing a directed cloning approach has been accomplished. The present application proposes to continue this search until a comprehensive inventory of all human serine proteases capable of interacting with APP-KI has been completed. This will enable establishment of the role that these proteases play in human disease. Related work is concerned with investigating the interactions of the Alzheimer's amyloid beta peptide and its fibrils with fibrin(ogen)-binding proteins. This group of investigators recently discovered that the amyloid beta peptide is a potent stimulator of human tissue plasminogen activator. This property of the beta peptide may in part explain the association of deposits of beta peptide in the cerebral vasculature (cerebral amyloid angiopathy) with hemorrhagic strokes, especially those that occur in conjunction with thrombolytic therapy. These findings also provide a basis for understanding the hemorrhagic phenotype of the genetic disease, hereditary cerebral hemorrhage with amyloidosis-Dutch type (HCHWA-D). In the work proposed in this application, a plan for mapping the functional epitopes on the amyloid beta peptide that account for its fibrin(ogen) mimicry, and the development of monoclonal antibodies that block the interaction of beta peptide with fibrin(ogen)-binding molecules were described. In addition, the design of amyloid beta peptide analogues that form soluble "minifibrils" amenable to structural analysis by high resolution methods was described. Finally, these investigators intend to explore whether synergistic interactions occur in the CNS between fibrin(ogen)-binding proteins and beta amyloid that may have relevance to Alzheimer's disease.

描述（改编自申请人摘要）本申请的长期目标是确定这三个通过分析APP的个体结构，功能域富含半胱氨酸的N-末端结构域已被表达并纯化至均质。优化表达生产适合X射线的材料需要结构晶体学和NMR。为了更好地了解生理 APP的行为--无论是在正常组织中还是在阿尔茨海默氏症的背景下疾病-试图确定APP结合蛋白在身体重点是分离潜在的蛋白酶，与APP Kunitz抑制剂（APP-KI）结构域。成功克隆一个这种利用定向克隆方法的候选蛋白酶已经完成了本申请提出继续该研究直到有一个全面的清单，所有人类丝氨酸蛋白酶能够与APP-KI的互动已经完成。这将有助于建立这些蛋白酶在人类疾病中的作用。相关的工作是调查的相互作用，阿尔茨海默氏淀粉样β肽及其纤维蛋白原结合纤维 proteins. 这组研究人员最近发现， β肽是人组织纤溶酶原激活物的有效刺激物。 β肽的这一特性可以部分解释脑血管中β肽的沉积（脑淀粉样蛋白血管病）与出血性中风，特别是那些发生在与溶栓治疗结合。这些发现也提供了依据为了了解遗传性疾病的出血表型，遗传性脑出血伴淀粉样变性-荷兰型（HCHWA-D）。在在这个应用程序中提出的工作，映射功能的计划，淀粉样蛋白β肽上的表位，解释其纤维蛋白（原）模仿，以及开发单克隆抗体， β肽与纤维蛋白（原）结合分子相互作用被介绍了此外，淀粉样β肽类似物的设计，形成可溶性的“微纤维”，适合于通过高分子聚合物进行结构分析。阐述了解决方法。最后，这些研究人员打算探索CNS中是否存在协同相互作用，纤维蛋白（原）结合蛋白和β淀粉样蛋白，可能与老年痴呆症