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（根据申请人的摘要改编）该应用的长期目标是确定这三个通过分析其个体的结构的尺寸结构功能域。富含半胱氨酸的N末端结构域已表达高水平并纯化为同质性。表达的优化生产适合X射线的材料需要构造晶体学和NMR。为了更好地了解生理应用的行为 - 在正常组织和阿尔茨海默氏症的背景下疾病 - 正在尝试识别应用程序结合蛋白的尝试身体。浓度是隔离相互作用的潜在蛋白酶使用App Kunitz抑制剂（APP-KI）域。成功克隆这种使用定向克隆方法的候选蛋白酶已经成就。本应用程序建议继续此搜索直到所有人类丝氨酸蛋白酶的全面清单与App-KI的互动已经完成。这将使建立这些蛋白酶在人类疾病中的作用。相关工作涉及调查阿尔茨海默氏症的淀粉样β肽及其原纤维与纤维蛋白（OGEN）结合蛋白质。这组研究人员最近发现淀粉样蛋白 β肽是人体组织纤溶酶原激活剂的有效刺激剂。 β肽的这种特性可以部分解释 β肽的沉积在大脑脉管系统中（脑淀粉样蛋白血管病）带有出血性中风，尤其是发生在与溶栓疗法的结合。这些发现也提供了基础为了了解遗传疾病的出血表型，遗传性脑出血，淀粉样蛋白 - 直属型（HCHWA-D）。在本应用程序中提出的工作是映射功能的计划解释其纤维蛋白（OGEN）的淀粉样β肽的表位模仿和开发单克隆抗体的开发 β肽与纤维蛋白（OGEN）结合分子的相互作用为描述。此外，淀粉样β肽类似物的设计形成可溶性的“微纤维”，可通过高的结构分析描述了解决方法。最后，这些调查人员打算探索是否在中枢神经系统中发生协同相互作用可能与阿尔茨海默氏病。