High Throughput Crystallization / Imaging System

高通量结晶/成像系统

• 批准号: 8448366
• 负责人: Lawrence J Delucas
• 金额: $ 59.4万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-15 至 2014-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8448366
• 关键词: Address; Adherence; Alabama; Anthrax disease; Area; Biological Process; Capsid; Communities; Complex; Comprehensive Cancer Center; Crystallization; Crystallography; Cystic Fibrosis; Disease; Faculty; Funding; Heat shock proteins; Image; Image Analysis; Influenza; Integrin Signaling Pathway; Malignant Neoplasms; Membrane Proteins; Molecular; Monitor; Nucleocapsid; Plague; Process; Production; Proteins; Research Project Grants; Resolution; Roentgen Rays; Students; System; Temperature; Tuberculosis; Tularemia; United States National Institutes of Health; Universities; Vesicular stomatitis Indiana virus; Viral; aqueous; base; biological research; cytokine; inhibitor/antagonist; instrumentation; macromolecule; member; programs; protein complex; protein structure; research study; screening; structural biology; three dimensional structure

DESCRIPTION (provided by applicant): In the past fifteen years it has become increasingly clear that atomic structural information via x-ray crystallography has accelerated and enhanced our understanding of the fundamental molecular mechanisms behind disease processes. For the majority of challenging aqueous and membrane proteins, the ability to rapidly determine three-dimensional structures has become more and more dependent on our ability to produce diffraction-quality three dimensional crystals of the target protein. This proposal addresses a critical need of the structural biology community at the University of Alabama at Birmingham (UAB): the availability of high throughput crystal imaging systems that can accommodate crystallization at the two most common temperatures, 20 ¿C and 4 ¿C. The UAB structural biology program includes 11 faculty-level crystallographers who conduct structural studies on macromolecules associated with a diverse scope of biological research areas, all of which involve medically relevant proteins and proteins involved in complicated biological processes. Some of the current projects include (1) structural studies on prokaryotic and eukaryotic membrane proteins, (2) bacterial adherence proteins, (3) viral capsid formation proteins, (4) proteins involved in the integrin signaling pathway, (5) vesicular stomatitis virus (VSV) nucleocapsid (6) heat shock proteins, (7) cytokines and (8) several protein structure-based discovery projects that involve developing inhibitors for protein targets associated with diseases such as cancer, cystic fibrosis, anthrax, plague, tularemia, viral influenza and tuberculosis. Each of these projects stands to benefit significantly from the proposed crystal imaging/analysis instrumentation which is expected to have a major impact on the progress of several NIH-funded research projects. Many of these projects involve highly labile/temperature sensitive proteins requiring 4 ¿C for purification and crystallization. In other cases (i.e. protein-protein complexes, protein-inhibitor complexes) crystals often display high mosaicity and/or poor diffracting resolution. However, many other projects previously crystallized, as well as many not yet crystallized, require trials at multiple temperatures since temperature is an important variabl in crystallization screening. For many of these challenging projects, the rate limiting step is the production of high-quality crystals. The addition of 4 ¿C and 20 ¿C high-throughput crystal imaging/analysis systems is requested to address the needs of eleven faculty level crystallographers and their pre-and postdoctoral level students. In addition, the requested systems will be incorporated into the Comprehensive Cancer Center's X-Ray Shared Facility thereby supporting any faculty member and/or students at the University of Alabama at Birmingham. The system's ability to remotely monitor a large number of crystallization experiments will enable utilization of this capability for several of the UAB faculty and students located in other buildings on campus.

描述（由申请人提供）：在过去的15年里，通过x射线晶体学获得的原子结构信息已经变得越来越清楚，它加速并增强了我们对疾病过程背后的基本分子机制的理解。对于大多数具有挑战性的水蛋白和膜蛋白，快速确定三维结构的能力越来越依赖于我们产生衍射质量的目标蛋白三维晶体的能力。该提案解决了阿拉巴马大学伯明翰分校（UAB）结构生物学社区的一个关键需求：高通量晶体成像系统的可用性，该系统可以在两种最常见的温度下进行结晶，20°C和4°C。UAB结构生物学项目包括11名教员级晶体学家，他们对与各种生物研究领域相关的大分子进行结构研究，所有这些研究都涉及与医学相关的蛋白质和参与复杂生物过程的蛋白质。目前的一些项目包括：(1)原核和真核膜蛋白的结构研究，(2)细菌粘附蛋白，(3)病毒衣壳形成蛋白，(4)整合素信号通路相关蛋白，(5)水泡性口炎病毒（VSV）核衣壳(6)热休克蛋白，(7)细胞因子和(8)一些基于蛋白质结构的发现项目，包括开发与癌症、囊性纤维化等疾病相关的蛋白质靶点抑制剂。炭疽、鼠疫、兔热病、病毒性流感和肺结核。这些项目中的每一个都将从拟议的晶体成像/分析仪器中受益匪浅，预计这将对nih资助的几个研究项目的进展产生重大影响。许多这些项目涉及高度不稳定/温度敏感的蛋白质，需要4°C进行纯化和结晶。在其他情况下（即蛋白质-蛋白质复合物，蛋白质-抑制剂复合物）晶体通常显示高嵌合性和/或低衍射分辨率。然而，由于温度是结晶筛选的重要变量，许多其他已经结晶的项目，以及许多尚未结晶的项目，都需要在多个温度下进行试验。对于许多具有挑战性的项目来说，限制速率的步骤是