500 MHz Solid-State NMR Spectrometer

说明(申请人提供)：需要一台500兆赫的固态核磁共振谱仪，以取代在购买的老化的360兆赫核磁共振仪。这台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，这些领域依赖于高分辨率固态核磁共振进行结构功能研究。第一个项目的重点是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。固体核磁共振方法正被用来确定视觉色素视紫红质的活性状态的结构，22-肾上腺素能受体与结合的激动剂和拮抗剂，以及CCR5与结合的HIV进入抑制剂。第二个项目(Erwin London)涉及膜筏和膜域的形成。完整的膜蛋白和外源性膜相关蛋白都与富含胆固醇的结构域结合，并可能起到核化作用。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缔合的机制。其中一个结构靶点是结构域形成蛋白小窝蛋白。小窝是小窝的蛋白质成分，小窝是负责膜内化的富含胆固醇的区域。第二个项目(斯图尔特·麦克劳克林)研究具有碱性残基和芳香族残基的跨膜和膜相关蛋白的结构和功能。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成员。受体酪氨酸激酶是细胞表膜受体，介导细胞的生长和分化，当通过突变或过度表达而被结构性激活时，与多种人类肿瘤有关。核磁共振结构研究的重点是受体活性是如何调节的。第四个项目(William Van Nostrand)的目标是阿尔茨海默病和脑淀粉样血管病(CAA)中涉及的淀粉样纤维的结构、形成和抑制。用固体核磁共振跟踪淀粉样多肽从单体到低聚物再到纤维的结构变化。重点是A242肽以及A240肽的荷兰和爱荷华州突变体。与公众健康相关：在过去的十年里，固态核磁共振光谱学已经成为一种有效的方法来确定细胞成分的高分辨率结构，这些结构不适用于结构生物学中的传统方法，即X射线结晶学和溶液核磁共振光谱学。这些结构包括与一系列神经退行性疾病(阿尔茨海默病、帕金森病、普里恩病)有关的淀粉样纤维，以及G蛋白偶联受体，这是目前市场上药物的主要靶标。所要求的仪器支持专注于这些生物医学目标的研究。

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.