Crystallization Robot for the Structural Biology Resource Center at Rockefeller

洛克菲勒结构生物学资源中心的结晶机器人

• 批准号: 7794108
• 负责人: Deena Abells Oren
• 金额: $ 14.73万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2011-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7794108
• 关键词: Biological; Budgets; Cell division; Computers; Crystallization; Custom; Dehydration; Deposition; Drops; Drug Design; Environment; Equipment; Eukaryotic Cell; Funding; Generations; Genetic Transcription; Growth; Hepatitis C virus; Housing; Lead; Manuals; Pathogenesis; Positioning Attribute; Price; Process; Proteins; Regulation; Request for Proposals; Research; Research Personnel; Research Support; Resources; Robot; Roentgen Rays; Sampling; Signal Transduction; Solutions; Spottings; Structure; System; Techniques; Time; Training; Universities; Virus Replication; Work; X-Ray Crystallography; discount; human disease; improved; instrument; macromolecule; nanolitre; protein-histidine kinase; public health relevance; research study; structural biology; success; three dimensional structure; wasting

DESCRIPTION (provided by applicant): This proposal requests funds to purchase a robot to set up crystallization experiments for the Structural Biology Resource Center at The Rockefeller University. The robot will aid researchers in obtaining diffraction quality crystals of biological macromolecules involved in bacterial transcription and its regulation, bacterial two-component histidine kinase signaling, bacterial pathogenesis, hepatitis C virus replication, and eukaryotic cell division. Obtaining crystals is often the rate-limiting step in X-ray structure determination. The robot will greatly increase the efficiency of crystallization trials by reducing the amount of time and sample required. This, in turn, will increase the success rate of NIH-funded research, and will increase utilization of the NIH-funded X-ray equipment already in the Resource Center. Due to the smaller quantities of materials required for the robot as opposed to manual setup, the robot will also allow extensive crystallization trials for projects that would otherwise not be possible. In addition, crystallization experiments will have a smaller impact on the environment by generating much less waste. The crystallization robot requested is the Screenmaker96+8 from Innovadyne - $145000. This robot can perform three distinct processes needed to set up crystallization experiments. Step 1: transfer of reservoir solutions from the sparse matrix stocks to the 96 positions in the crystallization tray; Step 2: deposition of nano-liter volumes of this reservoir onto the drop position and deposition of the protein nano-liter volume onto the same spot; Step 3: to refine the condition by making custom grids by varying two components in the lead condition to create new opportunities for improved crystal growth. Another very important feature of the Screenmaker96+8 is that the time required to place the drops (both the precipitant solution and the protein) is significantly less than other crystallization robots, thereby reducing dehydration time by at least half. The crystallization robot will be housed in the newly created Structural Biology Resource Center, supported by The Rockefeller University under the umbrella of the Research Support office. The Center will supervise the robot's use, train users and maintain the instrument in top working condition, and will be responsible for fair, relevant and efficient utilization of the robot. Additional parts needed: computer $2450, installation $4000, two nest chiller system $10450, for sensitive samples and extended warranty $32000. (Prices quoted here do not reflect 10% academic discount applied to total requested in budget, see price quote in Equipment.doc). PUBLIC HEALTH RELEVANCE: Determining the three-dimensional structure of biological macromolecules is key in understanding their function and in opening new venues for designing drugs and cures for human diseases. This can be accomplished through the technique known as X-ray crystallography, where the rate-limiting step is often the crystallization of the target molecule, a menial process that is labor intensive and time consuming. This proposal requests a robot that performs the steps involved in setting up crystallization trials of biologically important molecules, decreasing the labor 10 fold and decreasing the generation of waste while increasing the efficiency and scope of the experimental projects.

描述(由申请者提供)：这项提案要求购买一个机器人，为洛克菲勒大学的结构生物学资源中心建立结晶实验。该机器人将帮助研究人员获得生物大分子的衍射级晶体，这些大分子涉及细菌转录及其调控、细菌双组分组氨酸激酶信号转导、细菌发病机制、丙型肝炎病毒复制和真核细胞分裂。在X射线结构测定中，获得晶体通常是速度限制步骤。该机器人将通过减少所需的时间和样本量，大大提高结晶试验的效率。这反过来将提高NIH资助的研究的成功率，并将增加资源中心已有的NIH资助的X射线设备的利用率。由于与手动设置相比，机器人所需的材料数量较少，因此机器人还将允许对原本不可能实现的项目进行广泛的结晶试验。此外，结晶实验对环境的影响较小，因为产生的废物要少得多。所要求的结晶机器人是Innovadyne的Screenmaker96+8-145000美元。这个机器人可以执行建立结晶实验所需的三个不同的过程。步骤1：将储存液从稀疏的基质储存转移到结晶盘中的96个位置；步骤2：将该储存物的纳升体积沉积到滴落位置，并将蛋白质纳升体积沉积到相同的位置；步骤3：通过改变铅条件下的两种成分制作定制网格来改进条件，为改进晶体生长创造新的机会。ScreenMaker 96+8的另一个非常重要的特点是放置液滴(沉淀液和蛋白质)所需的时间明显少于其他结晶机器人，从而将脱水时间减少了至少一半。结晶机器人将安置在新成立的结构生物学资源中心，该中心由洛克菲勒大学在研究支持办公室的保护伞下提供支持。该中心将监督机器人的使用，培训用户并保持仪器处于最佳工作状态，并将负责公平、相关和有效地使用机器人。需要额外的配件：电脑2,450美元，安装4,000美元，两套冷冻机系统10450美元，用于敏感样品和延长保修期32000美元。(此处引用的价格不反映对预算中要求的总金额适用10%的学术折扣，请参阅Equipment.doc中的报价)。 与公共卫生相关：确定生物大分子的三维结构是了解它们的功能和为设计治疗人类疾病的药物和疗法开辟新途径的关键。这可以通过被称为X射线结晶学的技术来实现，在这种技术中，限速步骤通常是目标分子的结晶，这是一个劳动密集型和耗时的繁琐过程。这项提议要求机器人执行建立生物重要分子结晶试验所涉及的步骤，将劳动力减少10%，减少废物的产生，同时提高实验项目的效率和范围。

项目成果

X-ray structure of the mammalian GIRK2-βγ G-protein complex.

• DOI: 10.1038/nature12241
• 发表时间: 2013-06-13
• 期刊: Nature
• 影响因子: 64.8
• 作者: Whorton MR;MacKinnon R
• 通讯作者: MacKinnon R

Hooking She3p onto She2p for myosin-mediated cytoplasmic mRNA transport.

将 She3p 连接到 She2p 上以实现肌球蛋白介导的细胞质 mRNA 转运。

• DOI: 10.1073/pnas.1423194112
• 发表时间: 2015
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Singh,Nimisha;Blobel,Günter;Shi,Hang
• 通讯作者: Shi,Hang

DAXX envelops a histone H3.3-H4 dimer for H3.3-specific recognition.

• DOI: 10.1038/nature11608
• 发表时间: 2012-11-22
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Elsaesser, Simon J.;Huang, Hongda;Lewis, Peter W.;Chin, Jason W.;Allis, C. David;Patel, Dinshaw J.
• 通讯作者: Patel, Dinshaw J.