Crystallization robotics to support X-ray crystallography at MUSC

结晶机器人为 MUSC 的 X 射线晶体学提供支持

基本信息

批准号：
8052518
负责人：
Christopher Davies
金额：
$ 12.49万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2012-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): With the advent of improved methods for phasing, the rate-limiting step to determine the structure of a macromolecular using X-ray crystallography is most often crystallization. The goal of crystallization is to find the best combination of parameters to produce crystals of sufficient quality so they can be used to generate high resolution diffraction data. Such a search involves testing an almost limitless number of parameters, including protein concentration, type of buffer and its pH, nature and concentration of precipitant, temperature, and use of additives or ligands. Hence, "crystallization space" is immense and if searched by low-throughput methods where crystal screens are pipetted manually, the process is severely limited by the amount of biological material that can be purified. The application of dispensing robots can dramatically change this situation because these can pipette protein in nanoliter volumes and therefore allow a much greater sampling of conditions with less biological material. Trays can also be set up considerably faster than manual methods. Although MUSC has a developing and thriving program in X-ray crystallography, including shared resources for protein production, our ability to produce crystal structures is severely hampered by the lack of crystallization robotics. This application aims to address that deficiency requesting the Phoenix RE nanoliter dispensing robot from Rigaku Inc. This will not only enhance the productivity of existing crystallography projects at MUSC, but will also stimulate new activity involving a wider group of investigators who aspire to use X-ray crystallography and have been unable to do so for lack of crystals. The broad benefits will be (a) the ability to tackle more challenging and potentially rewarding projects, including membrane proteins, (b) enhancement of the research environment at MUSC due to a greater emphasis on collaborative mechanistic studies of important biomedical problems that can be published in the higher impact journals, (c) increased competitiveness for NIH funding, (d) improved training environment for graduate students and post-docs and, (e) further development of X-ray crystallography and of structural biology in general through recruitment of new faculty. PUBLIC HEALTH RELEVANCE: This proposal requests funds to purchase crystallization robotics to enhance the X-ray crystallography facility that supports biomedical research at the Medical University of South Carolina. X-ray crystallography is a method of determining the structural details of macromolecules whose biological functions may be significant in human health and disease. Precise knowledge of the structure and function of such macromolecules contributes to better understanding of normal and abnormal biological processes and development of improved therapies to treat or cure major illnesses such as cancer, infectious diseases, inflammatory diseases and Alzheimer's disease.

描述（由申请人提供）：随着改进的相位方法的出现，使用X射线晶体学确定大分子结构的速率限制步骤最常见于结晶。结晶的目的是找到参数的最佳组合，以产生足够质量的晶体，以便它们可用于生成高分辨率衍射数据。这样的搜索涉及测试几乎无限的参数，包括蛋白质浓度，缓冲液类型及其pH值，沉淀剂的性质和浓度，温度以及添加剂或配体的使用。因此，“结晶空间”是巨大的，如果通过低通量的方法进行晶体屏幕的手动移动方式进行搜索，则该过程受到可以纯化的生物材料量的严重限制。分配机器人的应用可以极大地改变这种情况，因为这些可以在纳米素体积中的移液管蛋白，因此可以用更少的生物学材料对条件进行更大的采样。托盘也可以比手动方法快得多。尽管MUSC在X射线晶体学方面具有开发和繁荣的计划，包括用于蛋白质生产的共享资源，但由于缺乏结晶机器人技术，我们生产晶体结构的能力受到了严重阻碍。该应用程序旨在解决要求Phoenix Re nanoliter从Rigaku Inc.中分配机器人的缺陷。这不仅会提高MUSC现有晶体学项目的生产力，而且还将刺激涉及更广泛的研究人员，这些研究人员渴望使用X射线晶体学，并且由于缺乏晶体而无法使用X射线晶体学。广泛的好处将是（a）解决包括膜蛋白在内的更具挑战性和潜在有意义的项目的能力，（b）由于更加强调有关重要的生物医学问题的协作机械性研究，因此可以在MUSC的研究环境增强，这些研究可以发表在较高的影响期刊上的重要生物医学问题；晶体学和结构生物学的总体招募新教师。公共卫生相关性：该提案要求资金购买结晶机器人技术，以增强支持南卡罗来纳州医科大学的生物医学研究的X射线晶体学设施。 X射线晶体学是一种确定生物学功能可能在人类健康和疾病中重要的大分子的结构细节的方法。对此类大分子的结构和功能的精确知识有助于更好地理解正常和异常的生物学过程，并开发改良的治疗或治愈主要疾病的疗法，例如癌症，传染病，炎症性疾病和阿尔茨海默氏病。