Structures of Protein Aggregates by Solid-State NMR

通过固态 NMR 分析蛋白质聚集体的结构

• 批准号: 7931186
• 负责人: Chad M Rienstra
• 金额: $ 18.56万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7931186
• 关键词: Abbreviations; Amyloid; Anisotropy; Argon; Behavior; Binding; Brain; Chad; Chemicals; Chemistry; Collaborations; Communities; Complex; Data Analyses; Data Set; Development; Diagnostic; Dimerization; Disease; Electron Microscopy; Event; Fluorescence; Human Resources; Kinetics; Knowledge; Label; Laboratories; Lecithin; Literature; Magic; Maintenance; Mass Spectrum Analysis; Measurement; Membrane; Methods; Micelles; Modeling; Modification; Molecular Conformation; Monitor; Mutation; N-terminal; NMR Spectroscopy; Oxygen; Parkinson Disease; Pathology; Phage Display; Phosphatidic Acid; Physiologic pulse; Physiological; Preparation; Proteins; Protocols documentation; Public Health; Regulation; Reporting; Research; Research Personnel; Research Project Grants; Resolution; Sampling; Sedimentation process; Side; Signal Transduction; Site; Sodium Dodecyl Sulfate; Solid; Solutions; Specificity; Spin Labels; Staging; Streptococcus IgG Fc-binding protein; Stress; Stretching; Structural Models; Structure; Surface; Synaptic Vesicles; Techniques; Technology; Testing; Thioflavin T; Variant; Vesicle; Wood material; alpha synuclein; base; design; dimer; early onset; endosulfine; globular protein; human disease; improved; in vitro Model; in vivo; instrumentation; interest; light scattering; mutant; neuron development; oxidation; planetary Atmosphere; programs; protein aggregate; protein aggregation; research study; scaffold; small molecule; solid state; solid state nuclear magnetic resonance; tool

Description: The long-term objective of this research project is to develop a detailed atomic-resolution structural and functional understanding of protein aggregation and fibrillation relevant to human disease, principally with studies of alpha-synuclein. Wild type alpha-synuclein and its mutants (A30P, E46K and A53T) associated with early-onset Parkinson's disease (PD) will be examined in several structural states, including protofibrils, fibrils and membrane-associatedcomplexes. We propose that the chemical details of these structures determine the natural physiological function of alpha-synuclein and, in the mutants or under environmental stress, contribute to PD pathology. Moreover, we propose that conversion among several structural states is an essential feature of synucleins; this structural plasticity is likely to be required for neuronal development and maintenance. To examine these structures, magic-angle spinning (MAS) solid- state NMR (SSNMR) experiments will be employed, elucidating details of covalent chemistry, conformation and dynamics that are inaccessible to other experimental techniques. Complementary analysis and characterization techniques will include kinetic measurements (by thioflavin T fluorescence, light scattering and sedimentation analysis), high-resolution mass spectrometry, atomic force and electron microscopy, solution NMR and phage display. We will leverage these technologies along with sample preparation and isotopic labeling strategies to improve our understanding of protein aggregation relevant to disease in the specific case of alpha-synuclein, including: (1) the fundamental biophysical principles of alpha-synuclein aggregation and membrane association, (2) specific structural information that will enable rational design strategies for small molecules that modulate alpha-synuclein function and/or serve as diagnostic tools for PD, and (3) elucidation of specific structural interactions between alpha-synuclein and other brain proteins. Relevance to public health: Alpha-synuclein is a central player in Parkinson's disease. The precise relationships between alpha-synuclein structure and this disease are not yet well understood. Our proposed studies aim to advance this fundamental knowledge, which will assist the broader research community in developing improved diagnostic tools and therapies for Parkinson's disease.

描述：本研究项目的长期目标是开发详细的原子分辨率