STRUCTURE DETERMINATION OF RPRP GENE PRODUCT

我们正在提炼叙利亚基因重组片段的结构 仓鼠PrP基因产物的三维异核核磁共振分析。这 已知的蛋白质存在于至少两种不同的 构象。一种主要为α螺旋的溶液形式 (PrPC)和另一种聚合测试表形式(PrP Scrapie)。这个 结构二态论本身就很有趣，因为它证明了 氨基酸序列可以编码不止一种结构，但更多 重要的是，该序列在人类病理学中具有重要意义，因为 它与普恩病毒疾病有关。普恩病毒可以引起绵羊的瘙痒病， 疯牛病，库鲁，克鲁兹菲尔德-雅各布， Gerstmann-Staussler-Scheinker病和致死性家族性失眠 人类。该项目对药物设计和 更基本的层面，是我们理解蛋白质折叠的能力。在……里面 在结构细化的过程中，我们经常使用MidasPlus进行检查 我们的计算结果，并显示核磁共振约束 这个节目就是一场闹剧。检查的交互过程 结构和添加约束严重依赖于图形 因为我们有如此多的光谱重叠。除了……之外 分子图形学，我们使用了核磁共振波谱分析和数据库 管理程序Spaky管理来自大约1000个质子的数据 共鸣和3000多个否定。

We are refining the structure of a recombinant fragment of syrian hamster PrP gene product using 3 dimensional heteronuclear NMR. This prion protein is known to exist in at least two different conformations. One solution form that is predominantly alpha helical (PrPc) and another, aggregated beta-sheet form (PrP Scrapie). The structural dimorphism in itself is interesting since it demonstrates that amino acid sequences can encode more than one structure, but more importantly, the sequence has significance in human pathology because it is involved in prion diseases. Prions can cause scrapie in sheep, mad cow disease, and kuru, Cruetzfeld- Jakob, Gerstmann-Staussler-Scheinker disease, and fatal familial insomnia in humans. This project is significant for both drug design, and, on a more fundamental level, our ability to understand protein folding. In the structure refinement process, we constantly use MidasPlus to check the results of our calculations, and display the NMR constraints with the program noeshow. The interactive process of examining the sructure and adding in restraints is heavily dependent on graphics since we have such a great deal of spectral overlap. In addition to molecular graphics, we use the NMR spectral analysis and database management program SPARKY to manage the data from about 1000 proton resonances and over 3000 NOES.