Multi-Nuclear, Multi-Dimensional Experiments for Biomolecular NMR

本研究致力于进一步发展核磁共振波谱作为一种测定溶液中较大生物分子的三维结构的方法。直到最近，稳定同位素的标记和三维和四维异核核磁共振的使用才使大于10 kDa的蛋白质在溶液中的结构确定成为可能。这里的溶液结构测定了一种较大的α螺旋蛋白。这些挑战构成了提案的主干，将通过使用异核交叉极化提高效率，通过使用多量子滤光器和更多维度来提高分辨率，并改进关键的多维核磁共振实验的分析。核磁共振波谱方法的最新进展使测定溶液中较小蛋白质的三维结构成为可能。三维溶液结构与了解其生物学功能有关，可以传达动态信息，在其他方法因缺乏晶体而失败的情况下，可以通过核磁共振获得。这项建议描述了开发更先进的核磁共振方法以帮助分析更大的蛋白质、核酸及其络合物的溶液核磁共振谱的研究。这项研究将以中等大小的蛋白质作为模型系统进行。这些新方法将使更大的蛋白质和核酸的核磁共振结构测定成为可能，从而有助于更好地了解这一更大类别的生物分子的结构和功能。

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.