Structure and conformational dynamics of alpha-synuclein

α-突触核蛋白的结构和构象动力学

• 批准号: 8467770
• 负责人: Quyen Quoc Hoang
• 金额: $ 18.82万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8467770
• 关键词: Appearance; Biological Process; Brain; Cells; Cessation of life; Complex; Development; Disease; Disease Progression; Drug Design; Elderly; Employee Strikes; Fluorescence; Goals; Human; Kinetics; Learning; Lewy Bodies; Lewy Body Dementia; Life; Liposomes; Membrane; Memory; Methods; Molecular; Molecular Conformation; Molecular Structure; Multiple System Atrophy; Muscle; Neurodegenerative Disorders; Neurologic; Nuclear Magnetic Resonance; Parkinson Disease; Pathogenesis; Population; Preparation; Procedures; Process; Proteins; Psyche structure; Rattus; Recombinants; Resistance; Resolution; Roentgen Rays; Solutions; Structure; Structure-Activity Relationship; Therapeutic; Time; alpha synuclein; base; combat; cost; disability; drug development; enzyme activity; fluorescence imaging; insight; medical schools; mutant; novel; programs; single molecule; synuclein; synucleinopathy; tool

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is a common neurological condition that progresses from subtle loss of muscular coordination to severe physical and mental disability and eventual death. There is no cure and no treatments to slow the disease progression. Currently there are ~1 million PD sufferers in the US, which costs about $25 billion USD per year. These numbers are expected to triple by 2050 as the population of the elderly is increasing rapidly. Thus, effective disease-modifying treatments are desperately needed. To contribute to the drug development effort, the goal of this project is to determine the structure o ¿-synuclein (¿-syn) to gain insights into its function and its mechanisms in the pathogenesis of Parkinson's disease, which are critical for rational drug design. The aggregation of ¿-syn leads to appearance of so-called Lewy bodies (LBs) as a defining pathological hallmark of Parkinson's disease. Similar aggregates of ¿-syn are also implicated in other neurodegenerative diseases, including multiple system atrophy (MSA), dementia with Lewy bodies (DLB), and others collectively known as synucleinopathies. ¿-Syn is highly abundant in the brain and it appears to be important for learning and memory; however, its precise function is still unknown. It has no known enzymatic activity, thus its unknown biological function depends on its physical structure; moreover, it causes disease via a gain-of-toxic function that is associated with its structural transformation into highly ordered fibrils. Thus, detailed structural information of ¿-syn is crucil for understanding its function and to gain insights into its transformation into a disease-causing entity. However, such structural details are currently unavailable mainly because, until recently, ¿-syn lacked a persistent structure in solution and was thought to be a natively unfolded protein, which is not amenable for structure determination. However, we have recently isolated an ordered tetrameric form of ¿-syn in solution. We did so by developing an expression and purification procedure aimed at preserving its native structure throughout the purification process. At about the same time, Dennis Selkoe's group at Harvard Medical School independently demonstrated that ¿-syn isolated from rat brains and living human cells are also natively folded and tetrameric with striking resemblance to our recombinant ¿-syn. Moreover, we further demonstrated that our ¿-syn preparation was resistant to aggregation, did not compromise liposome membranes, and non-toxic to cells, thus strongly supporting that the ordered tetrameric ¿-syn is physiologically relevant. Now that we have an ordered form of ¿-syn that is amenable to traditional biophysical tools, the specific aims of this proposal are; 1) To determine the atomic structure of ¿-syn; and 2) To determine the dynamics of its quaternary structure. We will do so by using a combination of X-ray crystallographic methods, nuclear magnetic resonance (NMR) methods, and single-molecular fluorescence methods.

描述（由申请人提供）：帕金森病（PD）是一种常见的神经系统疾病，从轻微的肌肉协调性丧失发展为严重的身体和精神残疾，直至最终死亡。目前尚无治愈方法，也没有治疗方法可以减缓疾病的进展。美国约有 100 万帕金森病患者，随着老年人口的迅速增加，预计到 2050 年这一数字将增加两倍，因此迫切需要有效的疾病缓解治疗。到药物开发工作，该项目的目标是确定 ¿ -突触核蛋白（¿-syn），以深入了解其在帕金森病发病机制中的功能和机制，这对于合理的药物设计至关重要。 -syn 导致所谓的路易体 (LB) 的出现，作为帕金森病的类似聚集体。 -syn 还与其他神经退行性疾病有关，包括多系统萎缩 (MSA)、路易体痴呆 (DLB) 以及其他统称为突触核蛋白病的疾病。 - Syn在大脑中含量很高，似乎对学习和记忆很重要；但其确切功能仍不清楚，因此其未知的生物功能取决于其物理结构；通过获得毒性功能来预防疾病，该功能与其结构转变为高度有序的原纤维有关，因此，¿ -syn 对于理解其功能并深入了解其向致病实体的转变至关重要，然而，目前尚无法获得此类结构细节，主要是因为直到最近，¿ -syn 在溶液中缺乏持久结构，被认为是一种天然未折叠的蛋白质，不适合结构测定。然而，我们最近分离出了 ¿ 的有序四聚体形式。我们通过开发一种表达和纯化程序来实现这一点，旨在在整个纯化过程中保留其天然结构。大约在同一时间，哈佛医学院的 Dennis Selkoe 团队独立证明了这一点。从大鼠大脑和活人细胞中分离出的 -syn 也具有天然折叠和四聚体，与我们的重组体惊人相似 ¿ -syn。此外，我们进一步证明了我们的 ¿ -syn 制剂具有抗聚集性，不会损害脂质体膜，并且对细胞无毒，因此有力地支持了有序四聚体 ¿ -syn 在生理上是相关的。现在我们有了 ¿ 的有序形式。 -syn 适合传统的生物物理工具，该提案的具体目标是 1) 确定 ¿ -syn；2) 为了确定其四级结构的动力学，我们将结合使用 X 射线晶体学方法、核磁共振 (NMR) 方法和单分子荧光方法。

项目成果

Potential pharmacological chaperones targeting cancer-associated MCL-1 and Parkinson disease-associated α-synuclein.

针对癌症相关 MCL-1 和帕金森病相关 α-突触核蛋白的潜在药理学伴侣。

• 发表时间: 2014-07-29
• 影响因子: 11.1
• 作者: Oh, Misook;Lee, Ji Hoon;Wang, Wei;Lee, Hui Sun;Lee, Woo Sirl;Burlak, Christopher;Im, Wonpil;Hoang, Quyen Q;Lim, Hyun
• 通讯作者: Lim, Hyun