500 MHz Solid-State NMR Spectrometer

500 MHz 固态核磁共振波谱仪

• 批准号: 7595314
• 负责人: STEVEN Owen SMITH
• 金额: $ 50万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595314
• 关键词: Adrenergic Receptor; Aging; Agonist; Alzheimer' s Disease; Amyloid; Amyloid Fibrils; Area; Binding; Biomedical Research; CCR5 gene; Caveolae; Caveolins; Cell surface; Cellular Structures; Cerebral Amyloid Angiopathy; Cholesterol; Drug Delivery Systems; Epidermal Growth Factor Receptor; ErbB Receptor Family Protein; Fluorescence Spectroscopy; Funding; G-Protein-Coupled Receptors; HIV Entry Inhibitors; Human; Integral Membrane Protein; Iowa; London; Marketing; Mediating; Membrane; Membrane Proteins; Methods; Mutation; NMR Spectroscopy; Neurodegenerative Disorders; Parkinson Disease; Peptides; Pharmacologic Substance; Prion Diseases; Proteins; Receptor Protein-Tyrosine Kinases; Research Support; Resolution; Retinal Pigments; Rhodopsin; Solutions; Structure; Universities; X-Ray Crystallography; cell growth; instrument; instrumentation; member; monomer; mutant; overexpression; public health relevance; receptor; solid state nuclear magnetic resonance; structural biology; tumor

DESCRIPTION (provided by applicant): A 500 MHz solid-state NMR spectrometer is requested in order to replace an aging 360 MHz NMR instrument that was purchased in 1989. This NMR instrument currently supports four areas of NIH-funded biomedical research at Stony Brook University that rely on high resolution solid-state NMR for structure-function studies. The first project focuses on the structure and mechanism of G protein-coupled receptors (Steven Smith). These receptors are important pharmaceutical targets. Solid-state NMR methods are being used to determine the structures of the active state of the visual pigment rhodopsin, the 22- adrenergic receptor with bound agonists and antagonists, and CCR5 with bound HIV entry inhibitors. The second project (Erwin London) concerns membrane rafts and membrane domain formation. Integral membrane proteins and extrinsic membrane-associated proteins both associate with and may serve to nucleate cholesterol-rich domains. NMR structural studies are used in combination with fluorescence spectroscopy to understand the mechanism of domain formation and protein association. One structural target is the domain forming protein, caveolin. Caveolin is the protein component of caveolae, cholesterol-rich domains responsible for membrane internalization. The second project (Stuart McLaughlin) deals with the structure and function transmembrane and membrane-associated proteins having clusters of basic and aromatic residues. One structural target is the epidermal growth factor (EGF) receptor, a member of the ErbB family of receptor tyrosine kinases. The receptor tyrosine kinases are cell-surface membrane receptors that mediate cell growth and differentiation, and are associated with a wide variety of human tumors when constitutively activated through mutation or overexpression. NMR structural studies are focused on how receptor activity is regulated. The fourth project (William Van Nostrand) targets the structure, formation and inhibition of amyloid fibrils involved in Alzheimer's disease and cerebral amyloid angiopathy (CAA). Solid-state NMR is used to follow the structural changes of amyloid A2 peptides from monomers to oligomers to fibrils. The focus is on the A242 peptide and the Dutch and Iowa mutants of the A240 peptide. PUBLIC HEALTH RELEVANCE: Over the past ten years, solid-state NMR spectroscopy has emerged as an effective method for determining the high-resolution structures of cellular components that are not amenable to the traditional approaches in structural biology, namely X-ray crystallography and solution NMR spectroscopy. These structures include amyloid fibrils associated with a wide range of neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, prion diseases) and G protein-coupled receptors, the major target of drugs currently on the market. The requested instrumentation supports research that focuses on these biomedical targets.

描述（由申请人提供）：需要一台500 MHz固态NMR光谱仪，以替换1989年购买的老化360 MHz NMR仪器。该NMR仪器目前支持斯托尼布鲁克大学NIH资助的生物医学研究的四个领域，这些领域依赖于高分辨率固态NMR进行结构-功能研究。第一个项目侧重于G蛋白偶联受体的结构和机制（Steven Smith）。这些受体是重要的药物靶标。固态核磁共振方法被用来确定视色素视紫红质，22-肾上腺素受体与绑定的激动剂和拮抗剂，和CCR 5与绑定的HIV进入抑制剂的活性状态的结构。第二个项目（欧文伦敦）涉及膜筏和膜结构域的形成。整合膜蛋白和外源性膜相关蛋白都与富含胆固醇的结构域相关，并可使富含胆固醇的结构域成核。NMR结构研究与荧光光谱结合使用，以了解结构域形成和蛋白质缔合的机制。一个结构靶标是结构域形成蛋白质，小窝蛋白。小窝蛋白是小窝的蛋白质组分，富含胆固醇的结构域负责膜内化。第二个项目（StuartMcLaughlin）涉及具有碱性和芳香残基簇的跨膜和膜相关蛋白的结构和功能。一种结构靶点是表皮生长因子（EGF）受体，其是受体酪氨酸激酶的ErbB家族的成员。受体酪氨酸激酶是介导细胞生长和分化的细胞表面膜受体，并且当通过突变或过表达组成型激活时与多种人类肿瘤相关。核磁共振结构研究的重点是受体活性是如何调节的。第四个项目（William货车Nostrand）的目标是参与阿尔茨海默病和脑淀粉样血管病（CAA）的淀粉样纤维的结构，形成和抑制。固态NMR用于跟踪淀粉样蛋白A2肽从单体到低聚物再到原纤维的结构变化。重点是A242肽和A240肽的荷兰和爱荷华州突变体。公共卫生关系：在过去的十年中，固态NMR光谱学已经成为一种有效的方法，用于确定细胞成分的高分辨率结构，这些结构不适合结构生物学中的传统方法，即X射线晶体学和溶液NMR光谱学。这些结构包括与广泛的神经退行性疾病（阿尔茨海默病、帕金森病、朊病毒病）和G蛋白偶联受体相关的淀粉样原纤维，G蛋白偶联受体是目前市场上药物的主要靶标。所要求的仪器支持侧重于这些生物医学目标的研究。