SOUTHEAST COLLABORATORY FOR STRUCTURAL GENOMICS

说明(摘自申请表)：本提案旨在开发、 集成并测试所有组件，以实现经济高效、 高通量原核生物结构基因组学研究 真核系统。目标有两个：(1)开发和测试 实验和计算策略，用于执行具有成本效益的 用X射线结晶学高通量测定蛋白质的结构 和核磁共振方法。(2)将这些策略应用于扫描一个人的整个基因组 快速生长的生物体。真核生物秀丽隐杆线虫和 一种与祖先有亲缘关系的原核生物微生物，具有小基因组， 其中具有代表性的基因组被选为金黄色葡萄球菌。作为人类的cDNA 从NCL赞助的基因组计划开始，他们将被包括在内 在这项研究中，最终目标是所有 人类基因组中的蛋白质。通过选择原核生物和真核生物 基因组，我们应该能够探索所有的障碍 对任何基因组的基因产物进行高通量结构测定。这个 Pilot Center下设三个工作组：蛋白质生产小组、核磁共振小组和 X射线结晶学小组。使用机器人方法，蛋白质生产 该集团计划每周为两种核磁共振提供大约80种蛋白质(每种蛋白质1-5毫克) 和X射线结晶学研究。虽然核磁共振小组已经开发出方法 这可能会导致高通量的方法，即X射线 结晶学小组在以下三个领域开发了创新方法 蛋白质结晶学。它们是：使用微克的蛋白质结晶 样本量，数据采集效率一年提高3至5倍 同步加速器位置，并在使用硫进行物相测定方面取得突破 异常散射信号。新的结晶过程将 对结构基因组学产生重大影响，因为几乎所有的蛋白质， 即使是那些可以用微克量表示的也将是候选 结晶学研究后者在以下方面具有重要意义 在可能的情况下，大规模结构确定的成本效益 一种解决蛋白质晶体结构的常规方法，而不需要 用硒原子代替硫，既省时又省钱。这 一项提案旨在整合这些开发和其他开发，以产生一个系统 用于经济有效的、高通量地确定原核生物 和真核系统。一旦完全开发，这个系统将戏剧性地 改变我们确定结构的速度。它将产生广泛的影响， 医学和生物技术在这个国家的发展。

DESCRIPTION (Taken from the Application): This proposal aims to develop, integrate, and test all of the constituents for carrying out cost-effective, high-throughput structural genomics research for both prokaryotic and eukaryotic systems. The objectives are two-fold: (1) To develop and test experimental and computational strategies for carrying out a cost-effective, high-throughput structural determination of proteins by X-ray crystallography and NMR methods. (2) To apply these strategies to scan the entire genome of an organism at a rapid pace. The eukaryotic organism, Caenorabditis elegans, and an ancestrally-related prokaryotic microorganism having a small genome, Pyrococcus furiosus, are selected as representative genomes. As human cDNAs from the NCl sponsored genome project become available, they will be included in the study with the ultimate goal being structural characterization of all proteins of the human genome. By selecting both prokaryotic and eukaryotic genomes we should be able to explore the full breadth of obstacles to high-throughput structure determination of gene products from any genome. The Pilot Center has three working groups: Protein Production Group, NMR Group, and X-ray Crystallography Group. Using a robotic approach, the protein production group plans to supply approximately 80 proteins/week (1-5 mg each) for both NMR and X-ray crystallographic studies. While the NMR Group has developed methods that potentially could lead to high-throughput methods, the X-ray crystallography group has developed innovative approaches in three areas of protein crystallography. They are: crystallization of proteins using microgram quantity of sample, increased data collection efficiency 3 to 5 times at a synchrotron site, and a breakthrough in phase determination using sulfur anomalous scattering signal. The new process of crystallization will significantly impact the structural genomics in that essentially all proteins, even those that can be expressed in microgram quantities will be candidates for crystallographic studies. The latter is significant in terms of cost-effectiveness for large-scale structure determination as it could allow one to solve a protein crystal structure routinely without the need of substituting the sulfur by selenium atom, saving both time and cost. This proposal aims to integrate these developments and others to produce a system for cost-effective, high-throughput structure determination on both prokaryotic and eukaryotic systems. Once fully developed, this system will dramatically change our speed of structure determination. It will have broad implications to medical science and biotechnology development in this nation.