Rutgers Helium Recovery System for High Field NMR

罗格斯高场核磁共振氦回收系统

• 批准号: 10170724
• 负责人: JEAN S BAUM
• 金额: $ 22.07万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10170724
• 关键词: Address; Amyloid; Architecture; Atomic Force Microscopy; Binding; Binding Sites; Biological; Biology; Biophysics; Cell physiology; Cells; Cellular Assay; Collagen; Collagen Fibril; Complex; Computing Methodologies; Connective Tissue Diseases; Cryoelectron Microscopy; Defect; Disease; Disease Progression; Extracellular Matrix; Health; Helium; Homeostasis; Hour; Inherited; Integrins; Intervention; Light; Link; Molecular; Molecular Conformation; Motion; Mutation; NMR Spectroscopy; Nature; Neurodegenerative Disorders; Osteogenesis Imperfecta; Parkinson Disease; Pathologic; Platelet aggregation; Play; Property; Proteins; Protocols documentation; Recovery; Role; Specificity; Structure; Surface; Surface Properties; System; Techniques; Therapeutic; Time; alpha synuclein; biological systems; biophysical tools; design; monomer; nanoscale; novel; novel therapeutic intervention; self assembly; solid state nuclear magnetic resonance; synucleinopathy

Abstract Protein self-assembly into structured fibrils plays both functional and dysfunctional roles in biology. We are investigating two classes of fibril forming proteins: collagen, a fibril that forms essential interactions with numerous proteins and extracellular matrix components for proper cellular function and α-synuclein (αS), an intrinsically disordered protein that self-assembles into oligomers and fibrils that are associated with debilitating synucleinopathies, such as Parkinson’s Disease. While in both cases, fibrils are often thought of as rigid, rather inert, entities, we have recently discovered conformational dynamics that profoundly impact their atomic- to-nano scale properties. Despite the importance of these fibrillar proteins, the molecular determinants of fibril- protein interactions and their impact in health and disease remain unanswered. Thus, the overarching objective of this proposal is to understand how molecular motions and surface properties modulate protein interactions at different assembly stages (monomer, oligomer, fibril), spatial extent (atomic to nanoscale), and temporal regime (picosecond to hours) to promote normal homeostasis or pathological disease states. The unifying theme of this proposal is that we are developing the key techniques and protocols necessary for fibril characterization that recognize the conformational plasticity and diverse interactions of these systems. We use a multifaceted approach integrating solution and solid-state nuclear magnetic resonance spectroscopy, atomic force microscopy, cryo-electron microscopy, computational methods, and link these to cellular experimentation. We are addressing the question of how collagen fibrils recognize their binding partners (we focus in particular on integrin, a key protein involved in platelet aggregation) despite the fact that many binding sites are hidden in the complex collagen fibril architecture. Beyond structure/function in healthy fibrils, we will investigate the impact of GlyX mutations in hereditary connective tissue disease such as Osteogenesis Imperfecta (brittle bone disease) and visualize for the first time how these defects impact on fibril assembly, structure and function. Although a very different biological system, we raise similar fibril interaction questions for αS: cell-to- cell propagation and templated seeding of endogenous αS monomers by fibrils increases the number of fibrils and is one of the primary factors in disease progression. This interaction mechanism is not understood and we will investigate this by first characterizing amyloid surfaces from the atomic to the nano-scale and then visualizing the interactions of the monomers with them. Results will shed light on the nature and specificity of collagen and αS interactions, the biophysical and biological impact of disease-affiliated collagen mutations, and mechanisms of pathological cell-to-cell propagation and seeding of αS aggregates. Elucidating novel interaction mechanisms will aid in design of new therapeutic strategies to rescue compromised collagen interactions or pathological αS aggregation in devastating neurodegenerative disease.

抽象的 蛋白质自组装成结构化的原纤维在生物学中扮演功能和功能失调的作用。我们是 研究两类的原纤维形成蛋白：胶原蛋白，一种与与 用于适当细胞功能的许多蛋白质和细胞外基质成分和α-突触核蛋白（αs） 本质上受干扰的蛋白质将自组合成寡聚物和纤维纤维的蛋白质 突触核酸，例如帕金森氏病。虽然在这两种情况下，原纤维通常都被认为是刚性的，但 相反，惰性，实体，我们最近发现了咨询动力，对他们的原子 - to-nano量表属性。尽管这些原纤维蛋白很重要，但原纤维的分子决定剂 蛋白质相互作用及其对健康和疾病的影响仍未得到解答。那是总体目标 该建议的是了解分子运动和表面特性如何调节蛋白质相互作用 在不同的组装阶段（单体，低聚物，原纤维），空间范围（原子至纳米级）和临时 政权（Picsecond to Hours）以促进正常的体内平衡或病理疾病状态。统一 该建议的主题是我们正在开发原纤维所需的关键技术和协议 表征认识到这些系统的构象可塑性和潜水员的相互作用。我们使用 多面方法整合溶液和固态核磁共振光谱，原子 力显微镜，冷冻电子显微镜，计算方法，并将其与细胞实验联系起来。 我们正在解决胶原纤维如何认识其约束伙伴的问题（我们特别关注） 在整联蛋白上，与血小板聚集有关的关键蛋白质）提出了一个事实，即许多绑定位点隐藏在 复杂的胶原纤维建筑。除了健康原纤维中的结构/功能之外，我们将研究 GlyX突变在颅联结组织疾病中的影响，例如成骨（脆性）（脆性 骨骼疾病）首次想象这些缺陷如何影响原纤维组件，结构和 功能。尽管一个非常不同的生物系统，我们为α提出了类似的原纤维相互作用问题：细胞到细胞 内源性αs单体的细胞传播和模板播种纤维增加了原纤维的数量 并且是疾病进展的主要因素之一。这种相互作用机制尚不理解，我们 将首先表征从原子到纳米尺度的淀粉样表面，然后 可视化单体与它们的相互作用。结果将阐明 胶原蛋白和αs相互作用，疾病相关胶原突变的生物物理和生物学影响， 病理细胞到细胞传播和αs聚集物的播种的机制。阐明小说 互动机制将有助于设计新的治疗策略来挽救受损的胶原蛋白 毁灭性神经退行性疾病中的相互作用或病理αS聚集。