Tools for High Throughput Structural Biology

高通量结构生物学工具

• 批准号: 7090132
• 负责人: GEORGE T DETITTA
• 金额: $ 311.8万
• 依托单位: HAUPTMAN-WOODWARD MEDICAL RESEARCH INST
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7090132
• 关键词: biomedical resource; cooperative study; fungal proteins; high throughput technology; protein structure; structural biology

DESCRIPTION (provided by applicant): We propose to assemble a technology development and structure determination pipeline for high throughput structural biology. Our emphasis will be on sample preparation for diffraction study. Our technology development efforts will center on (1) the expression and purification of small protein - protein complexes from yeast; (2) the expression and purification of transmembrane proteins from yeast; (3) the identification, expression and purification of antibodies that target transmembrane proteins from yeast, as aids to their crystallization; (4) the crystallization of protein complexes, transmembrane proteins, transmembrane protein - antibody complexes, and soluble proteins; and (5) the cryopreservation of protein crystals. All of the technology development efforts will be at, or eventually geared to, high throughput. Our structure determination pipeline will run at medium throughput, and will take projects emanating from the technology development pipeline through to structure solution, analysis and deposition. In addition we propose to serve as a structural biology research resource for the greater biological community, forming a partnership with biologists and structural biologists that will make the high throughput crystallization and cryopreservation pipeline available as a community resource. We currently serve the needs of more than 300 structural biologists who provide proteins for high throughput screening of chemical conditions that foster crystallization. We will extend that pipeline to include optimization and growth of frozen, mounted, diffraction-quality crystals ready for structural analysis. Access to the pipeline will rapidly extend the benefits of the technology development to a larger biological community.

描述（由申请人提供）：我们建议组装用于高通量结构生物学的技术开发和结构测定管道。我们的重点是衍射研究的样品制备。我们的技术开发工作将集中在（1）从酵母中表达和纯化小蛋白质-蛋白质复合物； (2)酵母跨膜蛋白的表达和纯化； (3) 鉴定、表达和纯化针对酵母跨膜蛋白的抗体，以帮助其结晶； (4)蛋白复合物、跨膜蛋白、跨膜蛋白-抗体复合物、可溶性蛋白的结晶； (5)蛋白质晶体的冷冻保存。所有技术开发工作都将实现或最终面向高吞吐量。我们的结构确定管道将以中等吞吐量运行，并将把来自技术开发管道的项目进行到结构解决、分析和沉积。 此外，我们建议作为更大的生物界的结构生物学研究资源，与生物学家和结构生物学家建立伙伴关系，这将使高通量结晶和冷冻保存管道作为社区资源可用。目前，我们满足 300 多名结构生物学家的需求，他们为促进结晶的化学条件的高通量筛选提供蛋白质。我们将扩展该管道，包括优化和生长冷冻、安装、衍射质量的晶体，为结构分析做好准备。进入管道将迅速将技术开发的好处扩展到更大的生物界。