Presenilin Biology and the Mechanisms of Alzheimer's Disease

早老素生物学和阿尔茨海默病的机制

• 批准号: 10454838
• 负责人: OKSANA BEREZOVSKA
• 金额: $ 169.37万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454838
• 关键词: Active Sites; Address; Alzheimer' s Disease; Amyloid beta-42; Amyloid beta-Protein; Animals; Aspartic Endopeptidases; Astrocytes; Benign; Binding; Binding Sites; Biochemical; Biochemistry; Biological; Biological Assay; Biological Models; Biology; Catalytic Domain; Cells; Cellular biology; Cerebrum; Chronic; Cloning; Collection; Complex; Development; Disease; Docking; Engineering; Enzyme-Linked Immunosorbent Assay; Enzymes; Event; Exocytosis; Fluorescence Resonance Energy Transfer; Future; G-substrate; Glutamates; Grant; Hospitals; Human; Impairment; Individual; Learning; Libraries; Life; Link; Massachusetts; Mediating; Methods; Microscopy; Modeling; Molecular Conformation; Monitor; Mutagenesis; Mutation; Nature; Neurons; Nuclear; Pathogenicity; Pathologic; Pathway interactions; Peptide Hydrolases; Peptides; Pharmaceutical Chemistry; Phenylalanine; Plasmids; Process; Program Research Project Grants; Protein Structure Initiative; Proteins; Proteolysis; Public Health; Reading; Reagent; Reporting; Research; Research Project Grants; Role; Signal Transduction; Site; Stem Cell Development; Structure; Symptoms; Synapses; Therapeutic; Therapeutic Studies; Time; United States National Institutes of Health; Ursidae Family; Variant; Vesicle; Wit; Woman; Work; abeta accumulation; alpha secretase; base; beta secretase; cellular imaging; experience; experimental study; fascinate; gamma secretase; imaging genetics; insight; interest; mutant; notch protein; novel; preclinical development; presenilin; prevent; programs; small molecule; stem cell biology; synaptotagmin I; uptake

SUMMARY: Since its cloning in 1995 and its identification (under this grant) as an unprecedented intramembrane aspartyl protease in 1999, Presenilin has been implicated in a remarkable array of signaling events in all metazoans. PS was discovered through research on Alzheimer’s disease, but it was soon shown to confer functions necessary for life, including as the protease that enables Notch nuclear signaling. Therefore, continuing to decipher the structure, functions, and protein and small-molecule regulators of PS is a priority for fundamental cell biology. At the same time, the invariant cerebral accumulation of amyloid b-protein (Ab) decades prior to symptoms has made PS/g-secretase a rational target for mechanistic and therapeutic study in AD. Despite its pleiotropic role in biology, the protease’s structure was only recently reported at 3.4Å, leaving many structure-function details unresolved, and small molecules that can safely and potently modulate its cleavage of APP are just entering human trials. For these reasons, three collaborators with deep experience in the study of Presenilin over 20 years wish to apply a range of methods in cell biology, biochemistry, cellular imaging, genetics, stem cell biology and medicinal chemistry to tackle some of the thorniest questions in PS/g- secretase biology. To wit, based on our new model of PS processivity (Bolduc et al., eLife 2016), can we insert many different FAD-causing PSI mutations into the protein and identify which residues contribute to the S1’- S2’-S3’ active-site pockets we recently found to mediate the tri-peptide cleavages? What is the biological mechanism of coordinated b- and g-secretase processing within a novel complex? How does a new PS protein interaction discovered here, namely with GLT-1, help modulate glutamate uptake by astrocytes and perhaps also change g-secretase function? Can one identify and validate GSMs that are sufficiently potent yet selective to chronically shift g-secretase cleavages from toxic Ab42/43 to protective Ab37/38 peptides? How can we learn more about the participation of PS/g-secretase in stem cell development in the CNS? Here, we propose numerous interrelated aims that incorporate three cross-cutting themes that unite our work. First, all 3 projects will build on a new explanatory mechanism of g-secretase processing discovered under this grant: that its processive tri-peptide cleavages are dictated by 3 pockets in the presenilin enzyme controlling the engagement of substrates with the catalytic site. Second is our strongly shared interest in small-molecule modulators of the PS/g-secretase complex. Each of our 3 projects includes aims that will examine GSMs to assess effects on the PS functions we are individually studying. A third cross-cutting theme comes from our shared use of a large library of PS1 and APP plasmids, sensitive ELISAs, and advanced microscopy reagents -- all developed under this grant. In sharing these approaches and reagents, our PPG is not a collection of marginally related aims but rather a highly integrated program in which we use variations on common themes & methods to address major unsolved questions about g-secretase, the RIP mechanism, and its safe modulation to treat and prevent AD.

摘要：自1995年克隆以来，它（在本赠款下）被确定为前所未有的 早老素在1999年被发现与一系列显著的信号转导有关， 所有后生动物的事件。PS是通过对阿尔茨海默病的研究发现的，但很快就被证明 赋予生命所必需的功能，包括作为使Notch核信号传导成为可能的蛋白酶。 因此，继续破译PS的结构、功能、蛋白质和小分子调节剂是一个重要的研究方向。 基础细胞生物学优先。与此同时，淀粉样b蛋白在大脑中的恒定积累 (Ab)在症状出现前几十年，PS/g-分泌酶已经成为机制和治疗的合理靶点。 学习AD。尽管它在生物学中具有多效性，但这种蛋白酶的结构直到最近才被报道为3.4kDa， 留下许多结构-功能细节未解决，小分子可以安全有效地调节 它对APP的切割才刚刚进入人体试验阶段。出于这些原因，三位经验丰富的合作者 在早老素的研究超过20年，希望将一系列方法应用于细胞生物学，生物化学，细胞 成像，遗传学，干细胞生物学和药物化学，以解决PS/g中一些最棘手的问题， 分泌酶生物学也就是说，基于我们的PS持续合成能力的新模型（Bolduc等人，eLife 2016），我们可以插入 许多不同的FAD引起的PSI突变的蛋白质，并确定哪些残基有助于S1 '- S2 '-S3'活性位点口袋，我们最近发现介导的三肽裂解？什么是生物学 协调B-和G-分泌酶加工的机制在一个新的复杂？一种新的PS蛋白 这里发现的相互作用，即与GLT-1的相互作用，有助于调节星形胶质细胞对谷氨酸的摄取， 也改变G-分泌酶的功能？人们能否识别和验证GSM是足够有效的，但有选择性 慢性转移G-分泌酶裂解从有毒的Ab 42/43保护Ab 37/38肽？我们怎么能 了解更多关于PS/g分泌酶参与CNS干细胞发育的信息？在这里，我们建议 许多相互关联的目标，其中包括三个贯穿各领域的主题，这些主题将我们的工作联系在一起。首先，所有3个项目 将建立在一个新的解释机制的g-分泌酶处理发现根据这项赠款：它的 进行性三肽裂解由早老素酶中的3个口袋控制 有催化位点的底物。第二，我们对小分子调节剂的强烈共同兴趣， PS/G-分泌酶复合物。我们的3个项目中的每一个都包括检查GSM以评估对 我们正在单独研究的PS功能。第三个跨领域主题来自我们共同使用的一个大型 PS1和APP质粒库、灵敏的ELISA和先进的显微镜试剂--所有这些都是在 这个grant。在分享这些方法和试剂时，我们的PPG不是一个边缘相关目标的集合， 相反，这是一个高度集成的计划，我们使用共同主题和方法的变化来解决主要问题。 关于g-分泌酶、RIP机制及其治疗和预防AD的安全调节的未解决问题。