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; 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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 合成囊泡相关的蛋白Alpha-核蛋白与零星和帕金森氏病通过其在Lewy体内以及几种导致疾病早期发作的遗传多态性与零星和家庭帕金森氏病有关。 α-核蛋白在溶液中本质上是非结构化的，但与脂质膜相关后，它主要对螺旋结构进行构象变化。该蛋白的功能构象显示为膜结合形式。由于亚基之间的近端很幸运，因此NMR很难研究该蛋白的全球折叠。此外，学习蛋白质 - 膜相互作用的结构方面一般并不是一件容易的事。 NMR的快速发展方法（例如Trosy和脉冲偶极ESR光谱法（PDS））有望应对测量从一小部分到数十个甚至数百纳米纳米范围的较大距离的挑战。对» - 突触核蛋白的不满PDS研究在确定（SDS）和裂磷脂胶束的背景下阐明了蛋白质的结构。结果表明，PDS提供了高分辨率，并且可以以合理的精度测量距离。这项研究揭示了存在两个膜结合的存在，这些膜被短连接器隔开。但是，获得的数据表明，螺旋螺旋之间的非螺旋断裂可能是由于胶束系统的空间限制而导致的。