Chaperone-Enabled studies of epigenetic regulation enzymes

表观遗传调控酶的伴侣蛋白研究

基本信息

批准号：
8730671
负责人：
ANTHONY A KOSSIAKOFF
金额：
$ 132.14万
依托单位：
UNIVERSITY OF CHICAGO
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：
2010-09-30 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8730671
关键词：
Affinity Animal Model Binding Biological Biological Assay Biological Process Biology Cataloging Catalogs Catalysis Cell Nucleus Cells Chicago Communities Complex Contracts Crystallization DNA Repair DNA biosynthesis Disease Elements Ensure Environment Enzymes Epigenetic Process Family Gene Expression Gene Silencing Generations Genetic Recombination Goals Histones Human In Vitro Individual Knowledge Laboratories Lead Life Lysine Management Information Systems Mediating Mediator of activation protein Methylation Modification Molecular Molecular Chaperones Molecular Conformation Morphologic artifacts Multiprotein Complexes Mutation Organism Performance Phage Display Post-Translational Protein Processing Principal Investigator Probability Production Property Protein Engineering Protein Structure Initiative Proteins RNA Processing Reagent Regulation Research Research Infrastructure Research Personnel Resources Set protein Signal Transduction Site Specificity Structure Structure-Activity Relationship Surface System Technology Time United States National Institutes of Health Universities base cell type cellular imaging design drug development enzyme activity enzyme substrate complex flexibility gene repression histone methyltransferase histone modification imprint in vivo inhibitor/antagonist meetings member operation paralogous gene programs protein complex protein protein interaction public health relevance receptor recombinational repair research study success

项目摘要

DESCRIPTION (provided by applicant): The overarching goal of this project is to elucidate the molecular mechanism governing the catalysis and regulation of histone modification enzymes. We will use our Chaperone-Enabled Biology and Structure (CEBS) technology platform to study an important, but challenging group of epigenetic regulating enzymes. The lysine-specific histone methyltransferases (HMT) and their complementary partners, lysine demethylases (KDM) function as key mediators of epigenetic signaling through their actions as "writers and erasers" of post-translational modifications on histone proteins. Numerous recent studies have highlighted the importance of lysine methylation of histones leading to direct impact on DNA replication, repair, recombination, gene silencing, imprinting and RNA processes making these enzymes potential key targets for drug development. However, progress in gaining fundamental knowledge about structure-function relationships governing their modes of operation has been slow because they are multidomain proteins and have been recalcitrant to both structural and functional analyses. To overcome the existing barriers, we will generate specialized reagents called "synthetic affinity binders" or sABs that will be used as chaperones for crystallization, as well as customized affinity reagents for cell biologically applications. To accomplish our objectives we have assembled a world-class team of investigators that will exploit sAB reagents for both structure determination and high level biological assays. A major emphasis of our approach is to identify and structurally/biochemically characterize the molecular complexes in which the HMTs and KDMs function by using sABs that stabilize and enhance crystallization of the complexes. Thus, CEBS effort will rely on close ties to the large high throughput centers for protein production and structure determination of individual enzymes in multiple conformational states, enzyme-substrate complexes and multiprotein complexes. A unique strength of our approach is that we will provide crystallization chaperones and the information on interaction partners to the high throughput centers to greatly increase the probability of success of structure determination.

描述（由申请人提供）：该项目的总体目标是阐明组蛋白修饰酶催化和调节的分子机制。我们将使用我们的伴侣分子生物学和结构（CEBS）技术平台来研究一组重要但具有挑战性的表观遗传调节酶。赖氨酸特异性组蛋白甲基转移酶 (HMT) 及其互补伙伴赖氨酸脱甲基酶 (KDM) 通过对组蛋白翻译后修饰发挥“书写者和擦除者”作用，充当表观遗传信号转导的关键介质。最近的许多研究强调了组蛋白赖氨酸甲基化的重要性，它对 DNA 复制、修复、重组、基因沉默、印记和 RNA 过程产生直接影响，使这些酶成为药物开发的潜在关键靶标。然而，在获得有关控制其操作模式的结构-功能关系的基础知识方面进展缓慢，因为它们是多域蛋白质并且难以进行结构和功能分析。为了克服现有的障碍，我们将生产称为“合成亲和力结合剂”或 sAB 的专用试剂，将其用作分子伴侣结晶，以及用于细胞生物学应用的定制亲和试剂。为了实现我们的目标，我们组建了一支世界一流的研究团队，将利用 sAB 试剂进行结构测定和高水平生物测定。我们方法的一个主要重点是通过使用稳定和增强复合物结晶的 sAB 来识别和结构/生物化学表征 HMT 和 KDM 在其中发挥作用的分子复合物。因此，CEBS 的工作将依赖于与大型高通量中心的密切联系，用于蛋白质生产和多种构象状态下的单个酶、酶-底物复合物和多蛋白复合物的结构测定。我们方法的独特优势在于，我们将向高通量中心提供结晶伴侣和相互作用伙伴的信息，以大大提高结构测定的成功概率。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

CDR-H3 diversity is not required for antigen recognition by synthetic antibodies.

DOI：
10.1016/j.jmb.2012.11.037
发表时间：
2013-02-22
期刊：
JOURNAL OF MOLECULAR BIOLOGY
影响因子：
5.6
作者：
Persson, Helena;Ye, Wei;Wernimont, Amy;Adams, Jarrett J.;Koide, Akiko;Koide, Shohei;Lam, Robert;Sidhu, Sachdev S.
通讯作者：
Sidhu, Sachdev S.

A New Versatile Immobilization Tag Based on the Ultra High Affinity and Reversibility of the Calmodulin-Calmodulin Binding Peptide Interaction.