CHEMISTRY AND BIOLOGY OF ALZHEIMER'S AMYLOID PROTEINS

描述：(改编自申请者摘要)长期目标 本研究旨在确定淀粉样蛋白在病理中所起的作用。 阿尔茨海默氏症(AD)。尽管许多人取得了进展 实验室，淀粉样蛋白微小蛋白的确切化学性质 其成分尚待阐明。据推测， 这些辅助的β淀粉样蛋白(AB)在 淀粉样蛋白沉积与阿尔茨海默病的发病机制。因此，这项研究 具体目的是提纯和确定初级结构和 主要和次要淀粉样蛋白的翻译后修饰 神经炎斑块、血管壁和弥漫性沉积物中的成分。 将利用超微结构免疫化学来确定 这些沉积物中少量淀粉样组分的位置。这个 神经炎斑块、血管淀粉样蛋白、弥漫性淀粉样蛋白的生物学效应 它们的分离成分将在神经元和神经胶质细胞上进行评估 培养中的细胞。 为了实现这些目标，将从AD脑中提纯淀粉样蛋白 以及来自血管的匀浆，以及最近开发的 包括过滤、等度蔗糖分离、 超速离心法、胶原酶水解法、洗涤剂溶解法和 不连续的蔗糖密度梯度。淀粉样蛋白的增溶作用 蛋白质通过变性、杂化和分散剂 然后使用多个高性能步骤进行纯化 高效液相色谱分析。将产生针对该病毒的抗血清 不同分离蛋白质组分及其在光和电子中的应用 显微免疫化学(胶体金)研究以确定其 原位定位和在纯化的淀粉样蛋白中的定位 衍生的酶和化学肽将通过以下方式进一步分级 高效液相色谱自动氨基酸分析和表征 微测序以及质谱分析。邮寄时间的厘定- 翻译修饰将通过气相色谱进行- 质谱学和二维高压电泳法。 阐明其化学性质和生物学作用 淀粉样蛋白将有助于理解 阿尔茨海默病的发病机制和病理生理学。这方面的知识也将有助于 在AD分子模型的开发中，并将有助于 实施治疗策略。

DESCRIPTION: (adapted from Applicant's Abstract) The long-term goal of this study is to establish the role that amyloid plays in the pathology of Alzheimer's Disease (AD). Despite the advances made by numerous laboratories, the exact chemical nature of the amyloid minor protein components still remains to be elucidated. It is hypothesized that these ancillary beta amyloid (AB) proteins are of great significance in amyloid deposition and in the pathogenesis of AD. Therefore, this study specifically aims to purify and determine the primary structures and post-translational modifications of the major and minor amyloid components from neuritic plaques vascular walls and diffuse deposits. Ultra structural immunochemistry will be utilized to ascertain the location of the minor amyloid components in these deposits. The biological effects of neuritic plaque, vascular amyloid, diffuse amyloid and their separated components will be assessed on neuronal and glial cells in culture. To achieve these goals, amyloid will be purified from AD brain homogenates as well as from blood vessels by recently developed protocols which include filtration, isocratic sucrose separations, ultracentrifugation, collagenase hydrolysis, detergent lysis and discontinuous sucrose density gradients. Solubilization of the amyloid proteins by denaturing, chaotropic and dispersing agents will be followed by their purification using multiple steps of high performance liquid chromatography (HPLC). Antisera will be generated against the different separated protein fractions and used in light and electron microscopic immunochemical (colloidal gold) studies to determine their location in situ and in purified amyloid.The relevant proteins and derived enzymic and chemical peptides will be further fractionated by HPLC and characterized by automatic amino acid analysis and microsequencing as well as by mass spectrometry. Determination of post- translational modifications will be carried out by gas chromatography- mass spectrometry and by two-dimensional high voltage electrophoresis. Elucidation of the chemical nature and of the biological actions of amyloid proteins will contribute to the understanding of the pathogenesis and pathophysiology of AD. This knowledge will also assist in the development of molecular models for AD and will aid in the implementation of therapeutic strategies.