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，其标识（根据这笔赠款）为前所未有的 1999年，膜内天冬氨酸蛋白酶在一系列显着的信号传导中隐含了presenilin 所有后生动物中的事件。 PS是通过关于阿尔茨海默氏病的研究发现的，但很快就显示了 对生命所必需的会议功能，包括作为实现Notch核信号的蛋白酶。 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 基本细胞生物学的优先级。同时，淀粉样B蛋白不变的大脑积累 （AB）症状前几十年使PS/G-分泌酶成为机械和治疗的合理靶标 AD研究。尽管蛋白酶的结构在生物学中起源于生物学作用，但直到最近才报告为3.4Å， 留下许多结构功能的细节未解决，以及可以安全和潜在调节的小分子 它对应用程序的裂解只是进入人类试验。由于这些原因，三位具有深厚经验的合作者 在20年以上的老年蛋白的研究中，希望在细胞生物学，生物化学，细胞中应用一系列方法 成像，遗传学，干细胞生物学和医学化学，以解决PS/g-中一些最棘手的问题 分泌酶生物学。基于我们的新的PS加工率模型（Bolduc等，Elife 2016），我们可以插入 许多不同的引起FAD引起的PSI突变进入蛋白质，并确定保留哪些有助于S1’- S2'-S3'活跃位点口袋我们最近发现可以介导三肽的切割？什么是生物学 新型复合物中协调的B-和G-分泌酶处理的机制？新的PS蛋白如何 在这里发现的相互作用，即与GLT-1，有助于调节星形胶质细胞的谷氨酸吸收 还会更改G-分泌酶功能吗？一个人可以识别和验证足够潜在但选择性的GSM 长期将G-分泌酶从毒性AB42/43转移以保护AB37/38肽？我们怎么能 了解有关PS/G-分泌酶参与中枢神经系统中干细胞发育的更多信息吗？在这里，我们建议 许多相互关联的目标结合了三个结合我们工作的跨切割主题。首先，所有三个项目 将建立在此赠款下发现的G-分泌酶处理的新剥夺机制上： 进程三肽裂解由控制参与的酶中的3个口袋决定 带有催化部位的底物。第二是我们对小分子调节器的强烈共同兴趣 PS/G-分泌酶复合物。我们的三个项目中的每个项目都包括将检查GSM的目标以评估对的影响 我们正在研究的PS函数。第三个横切主题来自我们对大型的共同使用 PS1和APP质粒，敏感ELISA和高级显微镜试剂的库 - 全部开发 这笔赠款。在共享这些方法和试剂时，我们的PPG不是一系列与少量相关的目标，而是 而是一个高度集成的程序，在该程序中，我们在该程序上使用常见主题和方法来解决专业 关于G-分泌酶，RIP机制及其安全调节和预防AD的尚未解决的问题。