High Throughput Protein Crystallography Using Zone Plate

描述(申请人提供)：确定与每个基因和功能相关的蛋白质结构对了解生命系统至关重要，并将在蛋白质工程、重组N-DA技术和基因治疗等领域产生重大影响。X射线蛋白质结晶学可能是确定蛋白质等大分子结构的最有用的技术，特别是在分子量超过30kD的情况下。XRadia提出了一种新的方法来提高基于实验室的蛋白质衍射系统的吞吐量和操作，使用波带片作为聚焦光学元件，并在紧凑的布局中使用明亮的微聚焦X射线源。在第一阶段，我们将论证拟议的概念的可行性，在现有的蛋白质衍射系统上使用现有的波带板，并证明在第二阶段开发一种紧凑的蛋白质结晶学系统是可能的，该系统的吞吐量至少比目前的系统高10倍，并且在安装和对准、在蛋白质晶体和检测器之间设置理想的聚焦位置方面易于操作。在第二阶段，我们计划制备在辐射收集角度和聚焦效率方面都有更好性能的波带片，并使用传统的微聚焦X射线源、波带片光学元件和CCD摄像机构建一个致密的蛋白质结晶学原型系统。这项工作的预期结果是高通量X射线衍射系统的工作原型，该系统也针对小分子晶体(约30毫米)进行了优化，这些晶体可以安装在台式计算机上，易于对准和操作，并且可以以相对较低的成本生产。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.