AUTOMATED CRYSTALLIZATION SCREENING OF REDOX ACTIVE PROTEINS

氧化还原活性蛋白的自动结晶筛选

• 批准号: 8360537
• 负责人: MARK A. WILSON
• 金额: $ 10.08万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360537
• 关键词: Arts; Biological; Biology; Collaborations; Communities; Crystallization; Drops; Funding; Grant; Liquid substance; National Center for Research Resources; Oxidation-Reduction; Persons; Principal Investigator; Proteins; Request for Proposals; Research; Research Infrastructure; Resources; Robot; Sampling; Screening procedure; Source; United States National Institutes of Health; Universities; Work; X ray diffraction analysis; X-Ray Diffraction; cost; instrument; nanolitre

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Advances in automated high throughput liquid handling have resulted in dramatically reduced sample requirement for macromolecular crystallization. Commercially available crystallization robots, such as the Gryphon produced by Art Robbins Inc, allow for nanoliter drop sizes and thus can screen for conditions that produce X-ray diffraction-quality crystals from as little as 1-2 mg of purified protein. Because the requirement for large amounts of sample is a major impediment to collaborative structural studies, reducing sample needs will expand the scope and quantity of collaborative structural work that can be pursued. This proposal requests funds to acquire the Gryphon crystallization robot. We propose that this acquisition will allow for greatly enhanced structural biological collaboration within the Redox Biology Center and the larger university community. Two RBC crystallographers, Drs. Joe Barycki and Mark Wilson, will be responsible for the instrument and for managing collaborations. Funds are also requested for a support person at the B.S. to M.S. level to maintain the instrument and to handle operational details of these collaborative efforts.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 自动化高通量液体处理的进步大大减少了大分子结晶所需的样品。商业上可用的结晶机器人，如Art Robbins Inc.生产的鹰头狮，允许纳升液滴大小，因此可以筛选出从低至1-2毫克的纯化蛋白质中产生X射线衍射质量晶体的条件。由于对大量样本的要求是协作结构研究的主要障碍，减少样本需求将扩大可进行的协作结构工作的范围和数量。这项提案要求获得资金来收购鹰头狮结晶机器人。我们认为，此次收购将极大地加强氧化还原生物学中心和更大的大学社区内的结构生物学合作。两位RBC结晶学家Joe Barycki博士和Mark Wilson博士将负责仪器和管理合作。还需要一名本科至硕士级别的支助人员来维护该文书，并处理这些协作工作的业务细节。