STRUCTURAL ASPECTS OF APP FUNCTION AND PATHOLOGY

描述(改编自申请人的摘要) 此应用程序的长期目标是确定这三个 通过对APP个体结构的分析得出APP的空间结构 功能域。富含半胱氨酸的N-末端结构域已被表达 在高水平，并纯化到均一。表达方式的优化 生产适用于X射线的材料将需要构造 结晶学和核磁共振。为了更好地了解生理学 APP的行为--无论是在正常组织中还是在阿尔茨海默病背景下 疾病--正在尝试确定APP结合蛋白在 尸体。重点放在分离潜在的相互作用的蛋白酶上。 与APP库尼茨抑制物(APP-KI)域。成功克隆了一个 利用定向克隆方法这种候选蛋白酶已经被 完成了。本申请建议继续进行这项搜索 直到对所有人类丝氨酸蛋白酶进行全面盘点 与APP-KI的互动已完成。这将使建立 这些蛋白酶在人类疾病中所起的作用。 相关工作涉及调查两个国家之间的相互作用 阿尔茨海默病淀粉样β蛋白及其与纤维蛋白原结合的纤维 蛋白质。这组研究人员最近发现，淀粉样蛋白 β-肽是一种强有力的人组织纤溶酶原激活剂。 β多肽的这一特性可以部分解释 脑血管系统(脑淀粉样蛋白)中的β-肽沉积 血管病)伴有出血性中风，尤指发生在 配合溶栓治疗。这些发现也提供了一个基础 为了了解这种遗传病的出血性表型， 遗传性脑出血伴淀粉样变性-荷兰型(HCHWA-D)。在……里面 本申请中提出的工作是绘制功能图的计划 淀粉样β蛋白多肽上的表位与其纤维蛋白(原) 拟态，以及开发出能够阻止 β-肽与纤维蛋白(原)结合分子的相互作用 描述。此外，淀粉样β蛋白多肽类似物的设计 形成适合于结构分析的可溶“微纤丝” 对拆分方法进行了描述。最后，这些调查人员打算 探讨中枢神经系统中是否存在协同作用 纤维蛋白(原)结合蛋白和β淀粉样蛋白可能与 阿尔茨海默氏症。

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.