STRUCTURAL ASPECTS OF ABPP FUNCTION AND PATHOLOGY

ABPP 功能和病理学的结构方面

• 批准号: 3123436
• 负责人: STEPHEN P ANDERSON
• 金额: $ 28.92万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3123436
• 关键词: X ray crystallography; alanine; amyloid proteins; cellular pathology; chemical binding; complementary DNA; esterase; fibrinolysis; genetic library; genetic manipulation; molecular cloning; molecular pathology; mutant; nuclear magnetic resonance spectroscopy; plasminogen activator; polymerase chain reaction; protein purification; protein structure function; serine proteinases; site directed mutagenesis

The long-term objective of the laboratory is a detailed characterization of the structure and the function of the Alzheimer's amyloid beta protein precursor (ABPP). Besides being implicated in Alzheimer's disease, this protein has recently been found in platelets, and a mutant version of the beta peptide itself has been linked to a hereditary cerebral hemorrhage, HCHWA-D. A description of the functional (or dysfunctional) behavior of ABPP in normal and pathological states might suggest new therapeutic approaches to the treatment of neurodegenerative diseases such as Aizheimers as well as cardiovascular disorders such as stroke. With these objectives in mind, the thrust of our proposed research is two-fold: a) to derive further high-resolution structural information about ABPP which might yield insights into function; and b) use the structural information that we already have to try to identify proteins in the brain and in the blood that specifically interact with ABPP or its subfragments. Our specific aims for the term of the grant are: 1. To begin a comprehensive study of the structure and function of ABPP by conducting a high resolution "alanine scan" of the N-terminal domain. 2. To produce large quantities of purified, recombinant ABPP N-terminal domain for structural studies by X-ray crystallography and multidimensional NMR. 3. To study the possible role of beta amyloid fibrils as stimulators of human tissue plasminogen activator. 4. To isolate molecular clones of cognate proteases or esterases that bind to the ABPP Kunitz domain. This work, if successful, should start to bring into focus the network of proteins in the body, both in the brain and in the blood, that interact with ABPP. This information will help elucidate the role ABPP plays both in healthy and in diseased tissue.

实验室的长期目标是详细表征 阿尔茨海默病β淀粉样蛋白的结构和功能 前体（ABPP）。 除了与阿尔茨海默病有关之外，这 最近在血小板中发现了蛋白质，并且该蛋白质的突变版本 β肽本身与遗传性脑出血有关， HCHWA-D。 对功能性（或功能失调）行为的描述 正常和病理状态下的 ABPP 可能会提出新的治疗方法 治疗神经退行性疾病的方法，例如 艾兹海默症以及中风等心血管疾病。 考虑到这些目标，我们提出的研究的主旨是 双重：a）获得进一步的高分辨率结构信息 ABPP 可能会产生对功能的见解； b) 使用结构 我们已经拥有的信息来尝试识别大脑中的蛋白质 以及在血液中与 ABPP 或其子片段发生特异性相互作用的物质。 我们在赠款期限内的具体目标是： 1. 开始全面研究ABPP的结构和功能 对 N 端结构域进行高分辨率“丙氨酸扫描”。 2. 生产大量纯化的重组ABPP N端 通过 X 射线晶体学和多维进行结构研究的领域 核磁共振。 3. 研究β淀粉样原纤维作为刺激物的可能作用 人体组织纤溶酶原激活剂。 4. 分离结合的同源蛋白酶或酯酶的分子克隆 到 ABPP Kunitz 域。 这项工作如果成功的话，应该会开始引起人们对网络的关注。 体内的蛋白质，包括大脑和血液中的蛋白质，相互作用 与ABPP。 此信息将有助于阐明 ABPP 所扮演的角色 在健康和患病组织中。