Reduced Dimensionality NMR Spectroscopy for Structural Genomics

本研究的重点是新型生物核磁共振技术的开发和应用，这些技术有望对结构基因组学以外的科学基础设施产生广泛的影响。通过支持“结构基因组学”中基因产物的功能分析，核磁共振有望在收获基因组测序项目的成果方面发挥重要作用。在这个“大科学”项目的框架下，蛋白质结构的大规模生产需要新的方法来确定经济有效的结构。本研究致力于进一步发展用于“结构基因组学”的降维三共振核磁共振实验。这种光谱学有望成为理想的选择，因为首先，它允许将核磁共振测量时间与信号检测的灵敏度要求相匹配，同时为自动光谱分析保留高数字分辨率；其次，与传统光谱学相比，它可以使用更小的核磁共振光谱集来分配蛋白质。目前的提案既需要基础研究，即生物核磁共振方法的开发，也需要针对核磁共振公园或为结构基因组学项目运营的联盟的商业效率的应用研究。研究的目标是开发一种协议，该协议允许在结构基因组企业中识别最小的核磁共振光谱集，以确定给定蛋白质的结构。考虑到RD核磁共振实验，当光谱仪时间的投资必须最小化时，这将允许一个保持高数字分辨率，因为它可以在结构基因组学项目的框架内预期。此外，为了在结构基因组学中的应用，将评估RD光谱对蛋白质自动共振分配的影响，并设计一个包含RD脉冲程序，NMR参数集和自动光谱分析作业文件的“RD NMR软件包”。此外，研究将侧重于改进RD实验的实施，包括一套允许测量稀液晶介质中残余偶极耦合的方案。这一发展开辟了一条途径，将它们的测量与共振分配合并，以及利用沿着多肽链的标量和残余偶极偶联的总和的变化来建立顺序连接。这项研究为学生在学术发展的各个阶段提供了绝佳的机会，并有望在整合研究和教育方面发挥重要作用。

SzyperskiMCB 0075773This research focusses on the development and application of novel Bio-NMR techniques which promise to have a broad imact for science infrastructure beyond use in structural genomics.NMR is expected to play an important role to harvest the fruits of genome sequencing projects by supporting the functional analysis of gene products in "structural genomics". The mass production of protein structures in the framework of this "big science" project requires new methodology for cost-effective structure determination. This research is dedicated to further develop reduced-dimensionality (RD) triple resonance NMR experiments for use in "structural genomics". Such spectroscopy promises to be ideally suited because it allows, firstly, matching of the NMR measurement time to sensitivity requirements for signal detection while retaining a high digital resolution for automated spectral analysis, and secondly, assigning proteins with a smaller set of NMR spectra when compared with conventional spectroscopy. The current proposal invokes both fundamental research, i.e., Bio-NMR methods development, and applied research targeting the commercial effeciency of NMR parks or consortia operated for structural genomics projects. Research will aim at the development of a protocol that allows identification of minimal sets of NMR spectra for structure determination of a given protein in a structural genomics enterprise. Considering RD NMR experiments, this will allow one retaining a high digital resolution when investments of spectrometer time have to be minimized, as it can be expected within the frameworkk of a structural genomics project. Moreover, for application in structural genomics, the impact of RD spectroscopy for the automatic resonance assignment of proteins will be evaluated, and an 'RD NMR package' comprising RD pulse programs, NMR parameter sets and job-files for automated spectral analysis will be devised. Moreover, research will focus on the implementation of improved RD experiments including a suite of schemes that allows measurement of residual dipolar couplings in dilute liquid crystalline media. This development opens an avenue to merge their measurement with the resonance assignment, as well as to make use of variations of sums of scalar and residual dipolar couplings along the polypeptide chain to establish sequential connectivities.This research offers outstanding opportunities for involving students at all stages in their academic development, and can be expected to play a valuable role in integrating research and education.