Crystallographic Automation

晶体学自动化

• 批准号: 7793204
• 负责人: T M Iverson
• 金额: $ 49万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2011-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7793204
• 关键词: Academic Medical Centers; Automation; Collaborations; Crystallization; Crystallography; Culicidae; Disease; Drops; Funding; Goals; Health; Human; Image; Laboratories; Lead; Lipids; Membrane Proteins; Minor; Molecular; Monitor; Nucleotides; Phase; Proteins; Reaction; Request for Proposals; Research Personnel; Robot; Robotics; Roentgen Rays; Sampling; Site; Structure; Techniques; Therapeutic; Time; Translational Research; Transportation; Universities; Work; design; experience; improved; insight; interest; macromolecule; nanolitre; outreach; protein structure; research study; structural biology; success

DESCRIPTION (provided by applicant): Over the past 10 years, it has become clear that imaging the atomic structures of macromolecules is important for both determining the molecular mechanisms of disease and to design therapeutics. This has lead to an increase in interest in determining crystal structures of proteins and nucleotides. For X-ray crystallographic studies, the identification of initial crystallization conditions is essential, but can take hundreds of trials for a soluble protein, and tens of thousands of trials for a membrane protein. This makes approaching these experiments daunting for researchers without prior experience in the field, and can inhibit collaborations between laboratories that use crystallography as a primary technique and laboratories focused on function. The proposal requests funds to enhance the crystallographic automation at Vanderbilt University through the purchase one robot for the crystallization of membrane proteins in cubic lipid phases (CLP) from Zinsserna (Northridge, CA) and one automated imager from Formulatrix (Waltham, MA) to document the time- course of crystallization reactions. The two goals of this purchase are: 1) to enhance the capabilities for crystallization of membrane proteins, and 2) to improve the capacity of the automated crystallization facility to allow outreach to researchers that do not specialize in structural biology. This proposal is currently endorsed by 14 laboratories at Vanderbilt University and the Vanderbilt University Medical Center, with 4 major users and 10 minor users. While these groups have had success using crystallographic automation consisting of a Mosquito nano-liter drop setter, there is no robot that can pipette the lipid phase within 1,000 miles. The expansion of the local crystallization robotic facility to include a module that pippettes the cubic lipid phase will have a broad positive impact on NIH-sponsored basic and translational research at Vanderbilt. Moreover, transportation of samples off-site to using imaging facilities will destroy these samples. Thus the on-campus ability to monitor an increased number of crystallization reactions will expand the number of laboratories that can use structural techniques as a part of their repertoire as they approach problems of human health. HEALTH RELEVANCE: The determination of structures of macromolecules can provide insight into the basic mechanisms of function and mis-function. To better approach difficult problems in crystallization, we propose to purchase a crystallization robot designed to work with membrane proteins, and an automated imager.

描述（由申请人提供）：在过去的10年中，人们已经清楚地认识到，大分子的原子结构成像对于确定疾病的分子机制和设计治疗方法都很重要。这导致了对确定蛋白质和核苷酸的晶体结构的兴趣增加。对于X射线晶体学研究，初始结晶条件的确定是必不可少的，但对于可溶性蛋白质可能需要数百次试验，对于膜蛋白质可能需要数万次试验。这使得没有该领域经验的研究人员难以进行这些实验，并且可能会抑制使用结晶学作为主要技术的实验室与专注于功能的实验室之间的合作。该提案要求提供资金，通过从Zinsserna（Northridge，CA）购买一台用于立方脂质相（CLP）中膜蛋白结晶的机器人和从Formulatrix（Waltham，MA）购买一台自动成像仪来记录结晶反应的时间过程，以提高范德比尔特大学的结晶自动化。此次收购的两个目标是：1）提高膜蛋白结晶的能力，2）提高自动结晶设施的能力，以便与不专门从事结构生物学的研究人员进行联系。该提案目前得到了范德比尔特大学和范德比尔特大学医学中心14个实验室的认可，其中有4个主要用户和10个次要用户。虽然这些研究小组已经成功地使用了由Moscow纳升液滴设置器组成的晶体自动化，但没有机器人可以在1,000英里内移液脂质相。本地结晶机器人设施的扩展，包括一个模块，移液立方脂质相将有一个广泛的积极影响，在范德比尔特NIH赞助的基础和转化研究。此外，将样本运至场外使用成像设施将破坏这些样本。因此，校园内监测越来越多的结晶反应的能力将扩大可以使用结构技术作为其解决人类健康问题的一部分的实验室数量。 健康相关性：测定大分子的结构可以深入了解功能和功能失调的基本机制。为了更好地解决结晶中的难题，我们建议购买一台用于膜蛋白的结晶机器人和一台自动成像仪。

项目成果

Structural Elucidation of Peptide Binding to KLHL-12, a Substrate Specific Adapter Protein in a Cul3-Ring E3 Ligase Complex.

肽与 KLHL-12（Cul3 环 E3 连接酶复合物中的底物特异性接头蛋白）结合的结构阐明。

• DOI: 10.1021/acs.biochem.9b01073
• 发表时间: 2020
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Zhao,Bin;Payne,WilliamG;Sai,Jiqing;Lu,Zhenwei;Olejniczak,EdwardT;Fesik,StephenW
• 通讯作者: Fesik,StephenW

A New Family of HEAT-Like Repeat Proteins Lacking a Critical Substrate Recognition Motif Present in Related DNA Glycosylases.

一个新的 HEAT 样重复蛋白家族，缺乏相关 DNA 糖基化酶中存在的关键底物识别基序。

• DOI: 10.1371/journal.pone.0127733
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Mullins,ElwoodA;Shi,Rongxin;Kotsch,LyleA;Eichman,BrandtF
• 通讯作者: Eichman,BrandtF