500 MHz Solid-State NMR Spectrometer

500 MHz 固态核磁共振波谱仪

基本信息

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
项目类别：
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&apos 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

项目摘要

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）：要求使用500 MHz固态NMR光谱仪，以取代1989年购买的360 MHz NMR仪器。目前，该NMR仪器目前支持NIH资助的生物医学研究的四个领域，该领域在Stony Brook University中依靠高分辨率固态NMR来依靠高分辨率固态NMR进行结构构造研究。第一个项目的重点是G蛋白偶联受体（Steven Smith）的结构和机理。这些受体是重要的药物靶标。固态NMR方法用于确定视觉色素视紫红质的活性状态，具有结合激动剂和拮抗剂的22-肾上腺素能受体以及具有结合HIV进入抑制剂的CCR5。第二个项目（Erwin London）涉及膜木筏和膜域的形成。整体膜蛋白和外在膜相关蛋白都与富含胆固醇的结构核定结合。 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关联的机制。一个结构靶标是形成蛋白质的结构靶标可可糖蛋白。小窝蛋白是富含胆固醇的结构域的蛋白质成分，负责膜内部化。第二个项目（Stuart McLaughlin）涉及具有基本和芳族残基簇的结构和功能跨膜和膜相关蛋白。一个结构靶标是表皮生长因子（EGF）受体，这是ERBB受体酪氨酸激酶家族的成员。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，当由突变或过表达组成性激活时，与各种各样的人类肿瘤相关。 NMR结构研究集中于受体活性如何调节。第四个项目（William van Nostrand）针对参与阿尔茨海默氏病和脑淀粉样血管病（CAA）的淀粉样蛋白原纤维的结构，形成和抑制。固态NMR用于遵循从单体到低聚物到原纤维的淀粉样蛋白A2肽的结构变化。重点放在A240肽的A242肽以及荷兰和爱荷华州突变体上。公共卫生相关性：在过去的十年中，固态NMR光谱已成为确定细胞成分的高分辨率结构的有效方法，这些结构不适合结构生物学的传统方法，即X射线晶体学和溶液NMR光谱。这些结构包括与广泛的神经退行性疾病（阿尔茨海默氏病，帕金森氏病，prion病）和G蛋白偶联受体有关的淀粉样蛋白原纤维，这是目前市场上市场上的主要药物。要求的仪器支持着重于这些生物医学目标的研究。