High Throughput Protein Crystallography Using Zone Plate

描述（由申请人提供）：确定与每个基因和功能相关的蛋白质结构对于理解生命系统至关重要，并将在蛋白质工程、重组DA技术和基因治疗等领域产生重大影响。x射线蛋白质晶体学可能是测定蛋白质等大分子结构最有用的技术，特别是对于分子量大于30kD的蛋白质。Xradia提出了一种新的方法来提高实验室蛋白质衍射系统的吞吐量和操作，采用带片作为聚焦光学和明亮的微聚焦x射线源在一个紧凑的布局。在第一阶段，我们将在现有的蛋白质衍射系统上使用可用的带板来证明所提出的概念的可行性，并表明在第二阶段有可能开发出紧凑的蛋白质晶体学系统，该系统的吞吐量至少比当前系统好10倍，并且在安装和对准方面易于操作，在蛋白质晶体和检测器之间设置理想的聚焦位置。在第二阶段，我们提出了在辐射收集角度和聚焦效率方面都有改进的带片，并使用传统的微聚焦x射线源、带片光学和CCD相机构建了一个原型紧凑的蛋白质晶体学系统。这项工作的预期结果是一个高通量x射线衍射系统的工作原型，该系统还针对小型（约30毫米）大分子晶体进行了优化，这些晶体可以安装在工作台上，易于对齐和操作，并且可以以相对适度的成本生产。该系统的商用版本可以极大地提高家庭实验室中晶体选择和数据收集的能力。

DESCRIPTION (provided by applicant): The determination of protein structure associated with each gene and functionality is crucial in understanding living systems and will have a significant impact in such areas as protein engineering, recombinant N DA technology and gene therapy. X-ray protein crystallography is probably the most useful technique to determine macromolecule structures such as proteins, especially for molecular weights above about 30kD. Xradia propose a new approach to improve the throughput and operation of lab-based protein diffraction system by the use of a zone plate as the focusing optic and a bright microfocus x-ray source in a compact layout. In Phase I, we will demonstrate the feasibility of the proposed the concept using available a zone plate on an existing protein diffraction system and show that it is will be possible to develop a compact protein crystallography system in Phase II that has a throughput at least 10X better than the current systems and ease of operation in terms of installation and alignment, setting of desirable focusing position between the protein crystal and the detector. In Phase II, we propose to fabricate zone plates with improved performance in both radiation collection angle and focusing efficiency, and construct a prototype compact protein crystallography system using a conventional microfocus x-ray source, zone plate optic, and CCD camera. The expected outcome of this effort is a working prototype of a high-throughput x-ray diffraction system that is also optimized for small (approximately 30 mm) macromolecular crystals that can fit on a benchtop and is easily aligned and operated, and that can be produced for a relatively modest cost. A commercially-available version of this system could dramatically increase the capabilities for crystal selection and data collection in the home lab.