Structural Biology and Biophysics of Alpha-Synuclein Fibrils by Solid-State NMR

通过固态核磁共振研究 α-突触核蛋白原纤维的结构生物学和生物物理学

基本信息

批准号：
10605819
负责人：
Collin Griffin Borcik
金额：
$ 6.91万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10605819
关键词：
Affect Affinity Amyloid Binding Binding Sites Biophysics Chad Characteristics Chemicals Collaborations Communities Complex Cryo-electron tomography Cryoelectron Microscopy Data Development Diagnosis Diagnostic Disease Environment Etiology Exhibits Faculty Fellowship Fluorescent Dyes Formulation Future Grant Imaging ligands In Vitro Infrastructure Institution International Joints Label Lansoprazole Length Lewy Body Dementia Ligand Binding Ligands Location Magnetic Resonance Measures Membrane Proteins Mentors Methodology Methods Molecular Conformation Motor Multiple System Atrophy NMR Spectroscopy National Research Service Awards Neurodegenerative Disorders Parkinson Disease Parkinson&apos s Dementia Pathogenicity Pathology Pathway interactions Patients Persons Phenotype Physiological Pittsburgh Compound-B Polymorph Positron-Emission Tomography Postdoctoral Fellow Preparation Property Protein Dynamics Public Health Publications Relaxation Research Residual state Resolution Risk Site Spectrum Analysis Structure Techniques Technology Temperature Testing Therapeutic Tissues Training Universities Variant Vertebral column Water Width Wisconsin aggregation pathway alpha synuclein clinical application cognitive function diagnostic tool disease phenotype experience experimental study graduate student improved in vivo insight meetings member molecular dynamics novel novel strategies preference protein aggregation protein structure small molecule solid state nuclear magnetic resonance structural biology synucleinopathy tool tool development

项目摘要

ABSTRACT Synucleinopathies are a major public health risk, with millions of people each year affected by Parkinson disease, Parkinson disease with dementia (PDD), Lewy Body dementia, and multiple system atrophy. These diseases impact both motor and cognitive function, for which there are no known cures and limited therapeutic options. It is therefore vital to determine the disease etiology, which is hypothesized to arise from the misfolding and aggregation of the protein α-synuclein into fibrils. The in vivo structural forms of these pathogenic fibrils will help to understand mechanisms of misfolding and aid in the development of imaging ligands with higher structure- specific binding. I will use solid state NMR (SSNMR) in combination with cryo-EM/ET to determine the structures of patient derived in vivo PDD fibrils. I will then investigate how in vivo fibril quaternary structure governs the stability and dynamics of mature diseased state fibrils and its effects on the aggregation pathway of α-synuclein fibrils. I will also investigate the interactions of imaging ligands to these fibrils to determine the structural motifs these compounds bind to by comparing binding site structure between in vitro and in vivo fibril preparations. Training plan: I have a considerable amount of research experience with SSNMR of membrane proteins, and I will add training in SSNMR methods required for structure determination of fibrils structures using novel approaches combining simulated annealing and molecular dynamics with cryo-electron microscopy (cryo-EM), cryo-electron tomography (cryo-ET) and SSNMR data. The National Magnetic Resonance Facility at Madison (NMRFAM) provides a world-leading environment for training with access to high field SSNMR spectrometers (600 to 900 MHz) and an ultra-high field (1.1 GHz) SSNMR spectrometer arriving in 2023. The infrastructure here will allow me to make new discoveries to both structure and dynamics of diseased fibrils and their interactions with imaging ligands. Furthermore, I will gain training in cryo-ET to obtain complementary data such as fibril width, twist, mass-per-unit length, and utilize cryo-EM to solve structures jointly with SSNMR to atomic resolution. These findings will be disseminated to the larger scientific community via publications and talks given at interdisciplinary meetings. My training will take place under Prof. Chad Rienstra, an internationally recognized leader in the field of biomolecular SSNMR, who has mentored dozens of graduate students and postdocs, many of whom are faculty members at top tier institutions. Environment: The University of Wisconsin-Madison is a highly ranked research university, with among the best environments available in the world for structural biology, with NMRFAM, the Cryo-EM Research Center, and the Center for High Throughput Computing. Prof. Katherine Henzler-Wildman, co-director of NMRFAM with Prof. Rienstra, has a strong background in insoluble protein structure and dynamics. Close collaboration with the cryo-EM group of Prof. Timothy Grant on fibril structure determination will be an added capability. The excellent research environment at UW-Madison with Prof. Rienstra will prepare me to be a future leader in structural biology and biophysics of complex biomolecules.

抽象的突触核蛋白病是主要的公共卫生风险，每年有数百万人受到帕金森病的影响，帕金森病伴痴呆（PDD）、路易体痴呆和多系统萎缩这些疾病。影响运动和认知功能，目前尚无已知的治疗方法，治疗选择也有限。因此，确定疾病病因至关重要，该病因是由错误折叠和错误折叠引起的。这些致病原纤维的体内结构形式将有助于蛋白质 α-突触核蛋白聚集成原纤维。了解错误折叠的机制并帮助开发具有更高结构的成像配体我将结合使用固态 NMR (SSNMR) 和冷冻电镜/电子断层扫描 (cryo-EM/ET) 来确定结构。然后我将研究体内原纤维四级结构如何控制。成熟病态原纤维的稳定性和动态及其对α-突触核蛋白聚集途径的影响我还将研究成像配体与这些原纤维的相互作用以确定结构基序。通过比较体外和体内原纤维制剂之间的结合位点结构来结合这些化合物。培训计划：我在膜蛋白的SSNMR方面有相当多的研究经验，并且我将增加使用新颖的方法测定原纤维结构所需的 SSNMR 方法的培训将模拟退火和分子动力学与冷冻电子显微镜（cryo-EM）相结合的方法，冷冻电子断层扫描 (cryo-ET) 和 SSNMR 数据。 (NMRFAM) 提供世界领先的培训环境，可使用高场 SSNMR 波谱仪（600 至 900 MHz）和超高场（1.1 GHz）SSNMR 波谱仪将于 2023 年抵达。在这里，我将能够对病变原纤维及其结构和动力学做出新的发现此外，我将接受冷冻电子断层扫描方面的培训，以获得诸如此类的补充数据。如原纤维宽度、扭曲、单位长度质量，并利用冷冻电镜与 SSNMR 联合求解原子结构这些发现将通过出版物和演讲传播给更大的科学界。我的培训将在国际公认的 Chad Rienstra 教授的指导下进行。生物分子SSNMR领域的领导者，指导了数十名研究生和博士后，其中许多其中是顶级机构的教员。环境：威斯康星大学麦迪逊分校是一所排名靠前的研究型大学，拥有世界上最好的结构生物学环境之一，与 NMRFAM、冷冻电镜研究中心和高通量计算中心 Katherine 教授合作。 Henzler-Wildman 是 NMRFAM 与 Rienstra 教授的联合主任，在不溶性蛋白质领域拥有深厚的背景与 Timothy Grant 教授的冷冻电镜团队在纤维结构方面的密切合作。威斯康辛大学麦迪逊分校的优秀研究环境和教授的决心将是一种额外的能力。 Rienstra 将使我成为复杂生物分子结构生物学和生物物理学领域未来的领导者。