Gene to Structure for $10K (GTS10K)

基因到结构仅需 1 万美元 (GTS10K)

• 批准号: 7652469
• 负责人: LANCE J STEWART
• 金额: $ 386.53万
• 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/7652469
• 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.

结构基因组学领域旨在通过对蛋白质家族代表成员的三维原子结构的实验阐明，结合对序列密切相关蛋白质模型的精确计算，来表征所有蛋白质的结构-功能关系。真核蛋白从基因到结构的过程充满了挑战，包括从cDNA文库中分离基因克隆容易出错，蛋白质表达和纯化的产量低，以及从结晶实验的建立到假定晶体的x射线衍射分析之间的时间浪费。针对这些挑战，我们提出发展一体化

项目成果

A Plug-Based Microfluidic System for Dispensing Lipidic Cubic Phase (LCP) Material Validated by Crystallizing Membrane Proteins in Lipidic Mesophases.

• DOI: 10.1007/s10404-009-0512-8
• 发表时间: 2010-06
• 期刊: MICROFLUIDICS AND NANOFLUIDICS
• 影响因子: 2.8
• 作者: Li, Liang;Fu, Qiang;Kors, Christopher A.;Stewart, Lance;Nollert, Peter;Laible, Philip D.;Ismagilov, Rustem F.
• 通讯作者: Ismagilov, Rustem F.

Gene Composer in a structural genomics environment.

结构基因组学环境中的 Gene Composer。

• DOI: 10.1107/s1744309111027424
• 发表时间: 2011
• 期刊: Acta crystallographica. Section F, Structural biology and crystallization communications
• 作者: Lorimer,Don;Raymond,Amy;Mixon,Mark;Burgin,Alex;Staker,Bart;Stewart,Lance
• 通讯作者: Stewart,Lance

Discovery of leukotriene A4 hydrolase inhibitors using metabolomics biased fragment crystallography.

• DOI: 10.1021/jm900259h
• 发表时间: 2009-08-13
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Davies DR;Mamat B;Magnusson OT;Christensen J;Haraldsson MH;Mishra R;Pease B;Hansen E;Singh J;Zembower D;Kim H;Kiselyov AS;Burgin AB;Gurney ME;Stewart LJ
• 通讯作者: Stewart LJ

Multiparameter screening on SlipChip used for nanoliter protein crystallization combining free interface diffusion and microbatch methods.

• DOI: 10.1021/ja908558m
• 发表时间: 2010-01-13
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Li, Liang;Du, Wenbin;Ismagilov, Rustem F.
• 通讯作者: Ismagilov, Rustem F.

Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer.

• DOI: 10.1186/1472-6750-9-37
• 发表时间: 2009-04-21
• 期刊: BMC biotechnology
• 影响因子: 3.5
• 作者: Raymond A;Lovell S;Lorimer D;Walchli J;Mixon M;Wallace E;Thompkins K;Archer K;Burgin A;Stewart L
• 通讯作者: Stewart L