MT VET COBRE PROJECT 4: ROLE OF COPPER IN PRION DISEASES

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。Pron病是哺乳动物的一种致命的神经退行性疾病，包括克雅氏病(人类)、瘙痒病(绵羊)、慢性消耗性疾病(麋鹿、鹿)和疯牛病(牛)。这些疾病的特征是错误折叠的PrP蛋白(PrP)在中枢神经系统中积累。众所周知，PrP(α-PrP)的正常形式在N-末端结构域内的几个位置与铜(II)结合。虽然金属配位和铜在这些位置上的相对亲和力的细节还没有完全了解，但一个共识是α-PrP能够结合~5个单核铜(II)离子。相比之下，人们对错误折叠的PrP中的铜环境知之甚少，尽管有迹象表明铜在这种和其他淀粉样蛋白疾病中发挥了作用。S在金属酶方面的专业知识为阐明错误折叠/感染性PrP中铜结合的细节及其在正常蛋白质转化为疾病状态中的作用提供了独特的视角。这些信息将有助于揭开Pron的繁殖和感染性的机制。McGuirl博士的实验室正在使用生物物理技术研究金属离子与各种异构体重组PrP的结合，包括荧光和电子顺磁共振(EPR)光谱、等温滴定热法(ITC)和电感耦合等离子体质谱(ICP-MS)。与一些已发表的报道相比，我们没有看到在α-PrP的C末端半部分存在高亲和力铜(II)结合位点的证据。对包括传染性PrP在内的其他亚型的研究正在进行中。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Prion diseases are fatal neurodegenerative diseases of mammals and include Creutzfeld-Jacob disease (humans), scrapie (sheep), chronic wasting disease (elk, deer), and mad cow disease (cattle). These illnesses are characterized by the accumulation of misfolded prion protein (PrP), in the central nervous system. It is well known that the normal form of PrP (alpha-PrP) binds copper(II) at several sites within the N-terminal domain. Although the details of metal coordination and relative affinities for copper at these sites are not completely understood, a consensus view is emerging that alpha-PrP is able to bind up ~5 mononuclear copper(II) ions. In contrast, little is known about the copper environments in misfolded PrP, despite the indications that copper plays a role in this and other amyloid diseases. The PI¿s expertise in metallo-enzymes provides a unique perspective to elucidate the details of copper binding in misfolded /infectious PrP, and its effect on the conversion of normal protein to the diseased state. This information will help unravel the mechanism of prion propagation and infectivity. Dr. McGuirl's lab is investigating metal ion binding to various isoforms of recombinant PrP using biophysical techniques, including fluorescence and electron paramagnetic resonance (EPR) spectroscopies, isothermal titration calorimetry (ITC), and inductively coupled plasma-mass spectrometry (ICP-MS). In contrast with some published reports, we see no evidence for a high affinity copper(II) binding site within the C-terminal half of alpha-PrP. Studies of the other isoforms, including infectious PrP, are ongoing.