OSU Crystallography Facility: Acquisition of Crystallization Robot

描述（由申请人提供）：在过去的十年中，俄亥俄州立大学的研究人员使用强大的x射线晶体学技术进行了各种高影响力的美国国立卫生研究院资助的研究，包括发现自然界中第22种遗传编码氨基酸，重要的RNA加工和DNA修复酶的结构，参与细菌群体感应的蛋白质，以及核激素受体的调节。来自俄亥俄州立大学三所不同学院的多名教员正在将这项技术用于他们研究项目的关键方面。一个拥有最先进的x射线衍射设备的共享设施已经启动并运行了四年。虽然在许多方面，俄亥俄州立大学在这一重要的研究领域处于优势地位，但一个主要的限制是缺乏自动化的结晶系统。x射线晶体学是一种非常强大的工具，但它需要将感兴趣的样品分子或复合物结晶，在大多数情况下，结晶是成功确定结构的主要瓶颈。该提案的目的是通过获得蚊子纳米体积自动移液器来减少俄亥俄州立大学x射线结构测定的瓶颈。虽然蚊子仪器比现有的方法更快、更准确地建立结晶实验，但其主要优点是每次实验使用的样品少了20倍。这使得研究人员可以对给定数量的样品测试20倍以上的结晶鸡尾酒溶液，这大大增加了找到合适结晶条件的机会。在通常情况下，当感兴趣的样本数量不足时，这可能是一个关键的优势。蚊子将允许俄亥俄州立大学的研究人员进行大约20倍的结晶实验，从而提高成功确定最佳结晶条件的机会。反过来，这将极大地有利于本提案中确定的主要和次要用户的广泛的美国国立卫生研究院资助的项目，以及许多其他美国国立卫生研究院资助的俄亥俄州立大学从事晶体学合作的研究人员。

DESCRIPTION (provided by applicant): In the past ten years investigators at Ohio State University have used the powerful technique of X-ray crystallography for a wide variety of high-impact NIH-funded research, including the discovery of the 22nd genetically encoded amino acid in nature, structures of important RNA processing and DNA repair enzymes, proteins involved in bacterial quorum sensing, and regulation of nuclear hormone receptors. Multiple faculty members from three different colleges at Ohio State are using this technology for key aspects of their research programs. A shared facility housing state-of-the-art X-ray diffraction equipment has been up and running for the last four years. While in many ways Ohio State is thus poised for strength in this important research area, a major limitation is the absence of an automated system for crystallization. X-ray crystallography is an extremely powerful tool, but it requires that the sample molecule or complex of interest be crystallized, and in most cases, crystallization is the major bottleneck along the path to a successful structure determination. The aim of this proposal is to reduce the bottleneck in X-ray structure determinations at Ohio State through the acquisition of a Mosquito nano-volume automated pipetter. While the Mosquito instrument sets up crystallization experiments considerably faster and more accurately than with existing methodologies, the principle advantage is that the instrument uses 20-fold less sample per experiment. This allows the researcher to test 20-fold more crystallization cocktail solutions for a given amount of sample, which dramatically increases the chances of hitting upon suitable crystallization conditions. This can be a critical advantage when the amount of the sample of interest is in short supply, as is often the case. The Mosquito will allow researchers at Ohio State to set up approximately 20-fold more crystallization experiments, and thus enhance the chance for success in identifying optimal crystallization conditions. This in turn will greatly benefit a wide-range of NIH-funded projects of the major and minor users identified in this proposal as well as many other NIH-funded researchers at Ohio State who are engaged in crystallographic collaborations. HEALTH RELEVANCE: Crystal structures of proteins provide critical insight into the functions of proteins, and how they interact with small molecules. This information can be of great assistance in the design of novel drugs and pharmaceuticals. The crystallization robot requested in this proposal will help to grow crystals faster and thereby accelerate the ability of our NIH funded investigators to address issues of fundamental relevance to human health.