PULSED DIPOLAR ESR STUDY ON MEMBRANE-BOUND ALPHA-SYNUCLEIN

膜结合 α-突触核蛋白的脉冲偶极 ESR 研究

• 批准号: 8364019
• 负责人: ELKA R GEORGIEVA
• 金额: $ 0.45万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364019
• 关键词: Address; Appearance; Binding; Data; Detergents; Disease; Electron Spin Resonance Spectroscopy; Funding; Genetic Polymorphism; Grant; Lead; Learning; Lewy Bodies; Link; Measures; Membrane; Membrane Lipids; Membrane Proteins; Methods; Micelles; National Center for Research Resources; Parkinson Disease; Physiologic pulse; Principal Investigator; Protein Conformation; Proteins; Research; Research Infrastructure; Resolution; Resources; Solutions; Source; Structure; Synaptic Vesicles; System; Technology; United States National Institutes of Health; alpha synuclein; cost; early onset; nanometer; protein folding; protein structure; synuclein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The synaptic vesicle-associated protein, alpha-synuclein, is linked to both sporadic and familial Parkinson's disease through its appearance in Lewy bodies and through several genetic polymorphisms that lead to early onset of disease. Alpha-synuclein is intrinsically unstructured in solution, but it undergoes conformational changes to a predominantly ¿-helical structure upon association with lipid membranes. The functional conformation of this protein was shown to be the membrane bound form. Global fold of this protein was difficult to study by NMR because of luck in the proximis between subunits. Moreover, learning the structural aspect of protein-membrane interactions is not an easy task in general. The rapidly developing methods of NMR, such as TROSY, and pulsed dipolar ESR spectroscopy (PDS) are poised to address the challenges of measuring a wide range of distances ranging from a small fraction to tens or even hundreds of nanometers. Resent PDS study on ¿-synuclein elucidated in considerable details the structure of the protein in the context of detergent (SDS) and lyophospholipid micelles. It was shown that PDS provides high resolution and distances can be measured with a reasonable accuracy. This study reveals the presence of two membrane bound ¿-helices separated by a short linker. However, obtained data suggest the non-helical break between the helices may result from the spatial confinement of the micelle system.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 突触囊泡相关蛋白，α-突触核蛋白，通过其在路易体中的出现和通过导致疾病早期发作的几种遗传多态性与散发性和家族性帕金森病相关联。α-突触核蛋白在溶液中本质上是非结构化的，但它在与脂质膜结合时经历构象变化，变为主要为半螺旋结构。该蛋白的功能构象被证明是膜结合形式。由于亚基之间的邻近性，该蛋白的整体折叠很难通过NMR研究。此外，了解蛋白质-膜相互作用的结构方面通常不是一件容易的任务。快速发展的NMR方法，如TROSY和脉冲偶极ESR光谱（PDS），有望解决测量从一小部分到数十甚至数百纳米的广泛距离的挑战。最近的PDS研究- 突触核蛋白阐明了相当详细的蛋白质的结构中的洗涤剂（SDS）和lyophospholipid胶束。结果表明，PDS提供了高分辨率和距离可以测量一个合理的精度。这项研究揭示了两个膜结合的<$-螺旋的存在下，由一个短的接头分开。然而，获得的数据表明，非螺旋之间的螺旋断裂可能是由于胶束系统的空间限制。