Crystallographic Automation

晶体学自动化

基本信息

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
项目类别：
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

项目摘要

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，很明显，成像大分子的原子结构对于确定疾病的分子机制和设计治疗剂都很重要。这导致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，必不可少的初始结晶条件的鉴定是必不可少的，但可以对可溶性蛋白进行数百次试验，并为膜蛋白进行数以万计的试验。这使得在没有事先经验的研究人员的情况下接近这些实验，并可以抑制使用晶体学作为主要技术和专注于功能的实验室的实验室之间的合作。该提案要求通过购买一个机器人来增强范德比尔特大学的晶体学自动化，以从Zinsserna（CAINDRIDGE，CA）中的立方脂质阶段（CLP）结晶，并从Formulatrix（MA，MA）进行自动化成像仪，以记录Crym-Artival cression for Crystallizatization creptimation creptallizatization reations的时间。这次购买的两个目标是：1）增强膜蛋白结晶的能力，以及2）提高自动结晶设施的能力，以允许向不专门研究结构生物学的研究人员推广。目前，范德比尔特大学（Vanderbilt University）和范德比尔特大学医学中心（Vanderbilt University Center）的14个实验室认可了该建议，其中有4位主要用户和10位未成年人使用。尽管这些群体使用晶体学自动化取得了成功，该晶体学自动化由蚊子纳米级落下器组成，但没有机器人可以在1,000英里内将脂质相移除。局部结晶机器人设施的扩展为包括一个模块，该模块将对范德比尔特的NIH赞助的基本和转化研究产生广泛的积极影响。此外，将样品运输到现场使用成像设施将破坏这些样品。因此，校园内监测增加的结晶反应的能力将扩大可以将结构技术作为其曲目的一部分的实验室数量，因为它们处理人类健康问题。健康相关性：确定大分子结构可以提供对功能和功能错误的基本机制的见解。为了更好地解决结晶中的困难问题，我们建议购买旨在与膜蛋白一起使用的结晶机器人和一个自动成像仪。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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。