Gene to Structure for $10K (GTS10K)

基因到结构仅需 1 万美元 (GTS10K)

• 批准号: 7256302
• 负责人: LANCE J STEWART
• 金额: $ 381.36万
• 依托单位: EMERALD BIOSTRUCTURES
• 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/7256302
• 关键词: Gene; Structure; 10K; GTS10K

Description: The field of structural genomics aims to characterize the structure-function relationships of all proteins by the experimental elucidation of the three dimensional atomic structures of representative members of protein families combined with the accurate computation of models for closely sequence related proteins. The path from gene to structure for eukaryotic proteins is riddled with challenges including the error-prone isolation of gene clones from cDNA libraries, poor yields in protein expression and purification, and time lost between the set-up of crystallization experiments and X-ray diffraction analysis of putative crystals. To address these challenges, we propose the development of integrated instruments, methods and software that will substantially increase the success rate while reducing the time, effort, materials, and cost required for protein structure determination. Among these new developments will be: (1) Rosetta-guided design and gene synthesis of expression optimized DNA sequences encoding proteins that are guided towards more facile purification and improved crystallogenesis; (2) nano-volume microfluidic crystallization devices that embody the major protein crystallization methods allowing extensive crystallization screening of precious protein samples and their complexes; and (3) a tunable in-house MAD X-ray source which will allow in situ room-temperature crystal X-ray diffraction screening and final anomalous dispersion diffraction data collection for rapid structure determination. Starting in year 3, these new technologies will be validated through the planned structure determination and public release of over 550 eukaryotic protein targets belonging to families with high relevance to cancer biology and therapy, including kinases, transcription factors, and metabolic enzymes. The dissemination of the proposed instrumentation, methods and software will allow researchers to pursue protein X-ray crystal structures at or below a total cost of $10,000 per eukaryotic protein structure.

描述：结构基因组学领域旨在通过实验性地阐明蛋白质家族的代表成员的三维原子结构，并结合了与密切序列相关蛋白的模型的准确计算，以实验性阐明所有蛋白质的结构 - 功能关系。从基因到结构的真核蛋白的路径充满了挑战，包括从cDNA文库中易于出现错误的基因克隆，蛋白质表达和纯化的产量差，以及在结晶实验的设置和X射线衍射之间丢失的时间推定晶体的分析。为了应对这些挑战，我们提出了综合的发展 仪器，方法和软件将大大提高成功率，同时减少蛋白质结构确定所需的时间，精力，材料和成本。在这些新的发展中，将是：（1）表达优化的DNA序列的Rosetta引导的设计和基因综合，编码蛋白质，这些蛋白被引导到更容易纯化和改善的结晶生成； （2）体现主要蛋白质结晶方法的纳米体积微流体结晶器件，允许对珍贵蛋白质样品及其复合物进行广泛的结晶筛选； （3）可调节的内部MAD X射线源，它将允许原位室温晶体X射线衍射筛选和最终异常分散衍射数据收集，以进行快速结构测定。从第3年开始，这些新技术将通过计划的结构确定和公开释放的550多个具有与癌症生物学和治疗高度相关的家族（包括激酶，转录因子和代谢酶）的家庭的公众释放。提议的仪器，方法和软件的传播将使研究人员以每一个真核蛋白质结构的总成本或总成本为10,000美元。