TOOLS FOR COMBINATORIAL MACROMOLECULAR CRYSTALLIZATION

三维X射线晶体结构信息在生物学研究中的重要性，从酶机制的阐明到抗艾滋病药物的设计，已导致科学界呼吁做出巨大努力，确定所有代表性蛋白质折叠的结构。在确定X射线晶体结构时，生长目标分子晶体的最关键步骤之一。不幸的是，大分子结晶方面的技术进步没有跟上大分子X射线结晶学的其他领域。我们提出了新型高密度结晶板的开发、原型生产和测试，以及用于收集和组织结晶结果数据的关系数据库软件。这些创新产品将把现代组合化学的空间、时间和省力的概念融入到大分子结晶中。这些硬件和软件工具的开发将产生一个集成包，旨在使任何分子生物学研究能够生长大分子晶体，这是使X射线结晶学成为其研究计划的常规部分的第一步。对于一个像确定所有代表性蛋白质折叠的三维结构这样雄心勃勃的项目来说，分子生物学各级研究人员的这种参与将是所需的努力规模。拟议的商业应用：拟议的研究具有巨大的商业价值，因为它将整合组合化学和X射线结晶学，这两项技术是现代药物发现中最强大的技术。第一阶段的成功将产生用户产出的研究工具，这些工具被设计为“开箱即用”，使任何分子生物学研究人员，无论以前的X射线结晶学经验如何，都能生长出其目标生物分子的X射线衍射质量的晶体。此外，这些第一阶段研究将使我们能够朝着我们的长期目标前进，即为需要高通量结晶筛选的大型项目实现生物大分子结晶的完全自动化。这种技术的发展将构成许多产品和专利的基础，例如自动结晶机器人。根据这一提议开发的产品的不断增长的市场非常大，因为它包括学术界和工业界的所有大分子X射线结晶学和分子生物学研究人员。

The importance of three-dimensional X-ray crystallographic structural information in biological research, ranging from the elucidation of enzyme mechanisms to anti-AIDS drug design, has led to the call for a massive effort in the scientific community to determine the structures of all the representative protein folds. In determining X-ray crystal structures, one of the most critical steps in growing crystals of the target molecule. Unfortunately, technological advancements in macromolecular crystallization have not kept pace with other areas of macromolecular X-ray crystallography. We propose the development, prototype production, and testing of novel high-density crystallization plates, and relational database software for collecting and organizing crystallization results data. These innovative products will incorporate the space, time, and effort-saving concepts of modern combinatorial chemistry into macromolecular crystallization. The development of these hardware and software tools will yield an integrated package designed to enable any molecular biology research to grow macromolecular crystals, the first step towards making X-ray crystallography a routine part of his or her research program. Such participation by researchers at all levels of molecular biology will be the scale of effort required for a project as ambitious as determining the three-dimensional structures of all the representative protein folds. PROPOSED COMMERCIAL APPLICATIONS: The proposed research has immense commercial value since it will integrate combinatorial chemistry and X-ray crystallography, two of the most powerful techniques in modern drug discovery. Success in Phase I will yield of user-yield research tools designed to be used "straight out of the box" to enable any molecular biology researcher, regardless of prior X-ray crystallography experience, to grow X-ray diffraction quality crystals of their target biomolecules. Furthermore, these Phase I studies will allow us to proceed toward our long-term goal of completely automating the crystallization of biological macromolecules for large scale projects requiring high-throughput crystallization screening. The development of such technology would form the basis for numerous products and patents, such as automated crystallization robots. The ever-growing market for the products developed under this proposal is very large, as it includes all macromolecular X-ray crystallography and molecular biology researchers in academia, as well as industry.