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.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 突触小泡相关蛋白，α-突触核蛋白，通过出现在路易小体和导致疾病早期发病的几个基因多态性，与散发性和家族性帕金森氏病有关。α-突触核蛋白在溶液中本质上是非结构化的，但当与脂膜结合时，它会经历构象变化，变成以螺旋为主的结构。该蛋白的功能构象为膜结合型。这种蛋白质的全球折叠很难用核磁共振来研究，因为幸运的是亚基之间的邻近。此外，学习蛋白质-膜相互作用的结构方面通常并不是一项容易的任务。快速发展的核磁共振方法，如TROSY和脉冲偶极ESR波谱(PDS)，正准备解决从小到几十甚至几百纳米的大范围距离测量的挑战。最近关于突触核蛋白的PDS研究相当详细地阐明了在洗涤剂(十二烷基硫酸钠)和溶磷脂胶束的背景下蛋白质的结构。结果表明，PDS具有较高的分辨率，能够以合理的精度进行距离测量。这项研究揭示了由一个短连接子分隔的两个膜结合的？螺旋的存在。然而，已获得的数据表明，螺旋之间的非螺旋断裂可能是由于胶束系统的空间限制造成的。