Multi-Nuclear, Multi-Dimensional Experiments for Biomolecular NMR

这项调查是关于进一步发展的 核磁共振光谱（NMR）作为一种方法， 确定较大生物分子的三维结构 在溶液中。 直到最近，用稳定的 同位素以及三维和四维异质结的应用 核磁共振可以测定蛋白质的结构， 溶液中10 kDa。 这里，溶液结构的确定 a larger较大alpha阿尔法helical螺旋proteins蛋白质. 这些挑战构成了 建议的主干，并将与改进接触 通过使用异向交叉极化， 通过使用多个量子滤波器和 更多的维度，并在分析 关键的多维核磁共振实验。 %%% 核磁共振方法的最新进展 光谱学使三种物质的测定成为可能- 溶液中较小蛋白质的三维结构。 三 维解结构与理解 它们的生物功能，可以传递动态信息， 在其他方法由于以下原因而失败的情况下，可以通过NMR获得 缺少水晶。 本提案介绍了对 开发更先进的核磁共振方法，以帮助分析 较大的蛋白质、核酸和 其复合物。 这些研究将在中等水平下进行- 大小的蛋白质作为模型系统。 新方法将使核磁共振 可用于较大蛋白质和核酸的结构测定 酸，从而有助于更好地了解结构 并在这一更大类的生物分子中发挥作用。

This investigation is concerned with the further development of Nuclear Magnetic Resonance Spectroscopy (NMR) as a method to determine the three-dimensional structures of larger biomolecules in solution. It is only very recently that labeling with stable isotopes and the use of three- and four- dimensional heteronuclear NMR has allowed the structure determination of proteins larger than 10 kDa in solution. Here the solution structure determination of a larger alpha helical proteins. These challenges form the backbone of the proposal and will be approached with improvements in efficiency by using heteronuclear cross polarization, with improvements in resolution by using multiple quantum filters and a larger number of dimensions and with improvements in analysis of key multi-dimensional NMR experiments. %%% Recent developments in the methods of Nuclear Magnetic Resonance Spectroscopy have made possible the determination of the three- dimensional structures of smaller proteins in solution. Three dimensional solution structures are relevant to the understanding of their biological functioning, can convey dynamic information and may be obtained by NMR in cases where other methods fail because of lack of crystals. This proposal describes investigations into the development of more advanced NMR methods to help analyze the solution NMR spectra of larger proteins, nucleic acids and complexes thereof. The studies will be carried out with a medium- sized protein as a model system. The new methods will make NMR structure determination possible for larger proteins and nucleic acids and thus contribute to the better understanding of structure and function in this larger class of biomolecules.