500 MHz Solid-State NMR Spectrometer

描述（由申请人提供）：需要一台500 MHz固态核磁共振光谱仪，以取代1989年购买的老化的360 MHz核磁共振仪器。该核磁共振仪器目前支持美国国立卫生研究院资助的石溪大学生物医学研究的四个领域，这些研究依赖于高分辨率固态核磁共振进行结构-功能研究。第一个项目主要研究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点。固体核磁共振方法被用来确定视觉色素紫红质、22-肾上腺素能受体结合激动剂和拮抗剂、CCR5结合HIV进入抑制剂的活性状态结构。第二个项目（Erwin London）涉及膜筏和膜域形成。整体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联，并可能参与成核。核磁共振结构研究结合荧光光谱来了解结构域形成和蛋白质结合的机制。一个结构靶点是结构域形成蛋白，洞穴蛋白。小窝蛋白是小窝的蛋白质成分，负责膜内化的富含胆固醇的结构域。第二个项目（Stuart McLaughlin）研究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。其中一个结构靶点是表皮生长因子（EGF）受体，它是ErbB受体酪氨酸激酶家族的一员。受体酪氨酸激酶是介导细胞生长和分化的细胞膜受体，当通过突变或过表达组成性激活时，与多种人类肿瘤有关。核磁共振结构研究的重点是如何调节受体活性。第四个项目（William Van Nostrand）的目标是参与阿尔茨海默病和脑淀粉样血管病（CAA）的淀粉样原纤维的结构、形成和抑制。固体核磁共振用于跟踪淀粉样蛋白A2肽从单体到低聚物到原纤维的结构变化。重点是A242肽和A240肽的荷兰和爱荷华突变体。公共卫生相关性：在过去十年中，固态核磁共振波谱已成为确定细胞成分高分辨率结构的有效方法，这是结构生物学中传统方法（即x射线晶体学和溶液核磁共振波谱学）无法适用的。这些结构包括与多种神经退行性疾病（阿尔茨海默病、帕金森病、朊病毒疾病）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点。所要求的仪器支持以这些生物医学目标为重点的研究。

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.