OSU Crystallography Facility: Acquisition of Crystallization Robot

描述(申请人提供)：在过去的十年里，俄亥俄州立大学的研究人员使用了强大的X射线结晶学技术进行了广泛的高影响力的NIH资助的研究，包括发现自然界中第22个遗传编码的氨基酸，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职员工正在将这项技术用于他们研究项目的关键方面。在过去的四年里，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。虽然俄亥俄州立大学在许多方面因此在这一重要研究领域具有实力，但一个主要限制是缺乏自动化的结晶系统。X射线结晶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络合物结晶，在大多数情况下，结晶是成功确定结构的主要瓶颈。这项提议的目的是通过购买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方法更快、更准确地建立结晶实验，但其主要优势是该仪器每次实验使用的样品少20倍。这使得研究人员可以针对给定数量的样品测试20倍以上的结晶鸡尾酒溶液，这极大地增加了找到合适结晶条件的机会。当感兴趣的样品数量供不应求时，这可能是一个关键的优势，这是经常发生的情况。蚊子将允许俄亥俄州立大学的研究人员建立大约20倍的结晶实验，从而增加成功确定最佳结晶条件的机会。这反过来又将极大地有利于本提案中确定的主要和次要用户的NIH资助的广泛项目，以及俄亥俄州立大学许多其他NIH资助的从事结晶学合作的研究人员。 与健康相关：蛋白质的晶体结构提供了对蛋白质功能以及它们如何与小分子相互作用的关键洞察。这些信息可以对新药和药物的设计有很大的帮助。这项提案中要求的结晶机器人将有助于更快地生长晶体，从而加快我们由NIH资助的研究人员解决与人类健康相关的根本问题的能力。

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.