Structure-inhibition of amyloid oligomers and fibrils

淀粉样蛋白寡聚体和原纤维的结构抑制

• 批准号: 7866473
• 负责人: STEVEN Owen SMITH
• 金额: $ 28.1万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7866473
• 关键词: AP40; Alzheimer' s Disease; Amino Acids; Amyloid; Amyloid Fibrils; Amyloid beta-Protein; Architecture; Atomic Force Microscopy; Biological Assay; C-terminal; Cells; Data; Deposition; Electron Microscopy; Elements; Face; Fluorescence; Gene Mutation; Glycine; Image; In Vitro; Inherited; Methods; Modeling; Molecular Sieve Chromatography; Molecular Structure; NMR Spectroscopy; Neurodegenerative Disorders; Neurons; Peptides; Phenylalanine; Protein Isoforms; Proteins; Resolution; Sampling; Side; Solutions; Structural Models; Structure; Surface; Tail; Testing; Thioflavin T; Touch sensation; Toxic effect; abeta oligomer; amyloid fibril formation; base; design; dimer; inhibitor/antagonist; instrument; monomer; neuron loss; novel; novel strategies; prevent; research study; solid state nuclear magnetic resonance; three dimensional structure

DESCRIPTION (provided by applicant): Amyloid deposits found in neurodegenerative diseases result from misfolding of cellular proteins. The challenge for developing specific inhibitors that block oligomer or fibril formation is that there are no high- resolution molecular structures that can guide the design. The proposal has three specific aims. The first aim is to use high-resolution solid-state NMR in combination with atomic force microscopy (AFM) to refine the structures that are emerging of Abeta oligomers and fibrils. The second aim is to design inhibitors based on the structural data from Aim 1 and to test their ability in vitro to block oligomer or fibril formation. The third aim is to assay the ability of these inhibitors to block the toxicity of Abeta oligomers and fibrils on neuronal cells. A new approach has been developed for obtaining high-resolution AFM images of Abeta soluble oligomers and fibrils in solution. The method takes advantage of a novel AFM controller that provides resolution in 'single touch' AFM experiments that surpasses the resolution currently available using commercial instruments. High resolution AFM of solution samples will allow us to follow the formation of Abeta oligomers, protofibrils and fibrils, and determine how designed inhibitors prevent fibrillization. We have developed structural models for the Abeta42 monomer, dimer, protofibril and fibril based on preliminary results from high resolution AFM and solid-state NMR. The structures show that when the b-strands have a parallel orientation and the amino acids are in-register with one another, the surface of the b-sheet has pronounced ridges and grooves. This architecture provides the key elements for the rational design of inhibitors to prevent fibril formation. Our template inhibitor peptide based on a rational design approach has the sequence GxFxGxF, where the bulky phenylalanine side chains of the inhibitor are predicted to pack against the glycines in the GxxxG motif of the amyloidogenic peptide. We will test the ability of the designed peptides to disrupt the formation of oligomers and fibrils by thioflavin T fluorescence, size exclusion chromatography, electron microscopy, AFM and solid-state NMR. We will also test the ability of our designed inhibitors to protect neurons from cell death induced by amyloid fibrils. We will focus on the Abeta42 peptide because of its higher ability to form aggregates than the shorter isoforms. Moreover, most gene mutations that are associated with the inherited forms of Alzheimer's disease result in an increase in the ratio of Abeta42 over Abeta40.

描述（由申请人提供）：在神经退行性疾病中发现的淀粉样蛋白沉积是细胞蛋白错误折叠的结果。开发阻断低聚物或纤维形成的特异性抑制剂的挑战在于没有高分辨率的分子结构可以指导设计。该提案有三个具体目标。第一个目标是使用高分辨率固态核磁共振结合原子力显微镜（AFM）来细化正在出现的β低聚物和原纤维的结构。第二个目标是根据aim 1的结构数据设计抑制剂，并在体外测试它们阻断低聚物或原纤维形成的能力。第三个目的是测定这些抑制剂阻断β低聚物和原纤维对神经元细胞的毒性的能力。