Tools for High Throughput Structural Biology

高通量结构生物学工具

• 批准号: 6960831
• 负责人: GEORGE T DETITTA
• 金额: $ 370万
• 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/6960831
• 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）：我们建议组装高通量结构生物学的技术开发和结构确定管道。我们的重点是用于衍射研究的样本准备。我们的技术开发工作将以（1）酵母的小蛋白质蛋白质复合物的表达和纯化为中心； （2）酵母跨膜蛋白的表达和纯化； （3）鉴定，表达和纯化的抗体靶向酵母的跨膜蛋白，以帮助其结晶； （4）蛋白质复合物，跨膜蛋白，跨膜蛋白 - 抗体复合物和可溶性蛋白的结晶； （5）蛋白质晶体的冷冻保存。所有技术开发工作都将达到或最终达到高吞吐量。我们的结构确定管道将在中等吞吐量下运行，并将将从技术开发管道到结构解决方案，分析和沉积的项目进行。 此外，我们建议为更大的生物界提供结构生物学研究资源，与生物学家和结构生物学家建立合作伙伴关系，以使高通量结晶和冷冻保存管道可作为社区资源可用。目前，我们满足了300多家结构生物学家的需求，这些生物学家为促进结晶的化学条件的高吞吐量筛查提供了蛋白质。我们将扩展该管道以包括对结构分析的冷冻，安装，衍射质量的晶体的优化和生长。进入管道将迅速将技术开发的好处扩大到更大的生物界。