Chaperone-Enabled studies of epigenetic regulation enzymes

表观遗传调控酶的伴侣蛋白研究

• 批准号: 8152115
• 负责人: ANTHONY A KOSSIAKOFF
• 金额: $ 130.25万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 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/8152115
• 关键词: Chaperone; Enabled; studies; epigenetic; regulation

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. PUBLIC HEALTH RELEVANCE: Lysine-specific histone methyltransferases and histone demethylases are critical enzymes in the epigenetic regulation of DNA replication and repair, as well as recombination and gene expression and repression. Mutations in these enzymes have been implicated in a number of diseases. These studies will provide the biological rationale and structural basis for drug development.

描述（由申请人提供）：本项目的总体目标是阐明控制组蛋白修饰酶的催化和调节的分子机制。我们将使用我们的伴侣生物学和结构（CEBS）技术平台来研究一个重要的，但具有挑战性的表观遗传调节酶组。赖氨酸特异性组蛋白甲基转移酶（HMT）及其互补伙伴赖氨酸去甲基化酶（KDM）作为表观遗传信号的关键介质，通过它们作为组蛋白翻译后修饰的“书写者和擦除者”发挥作用。最近的许多研究都强调了组蛋白赖氨酸甲基化的重要性，它直接影响DNA的复制、修复、重组、基因沉默、印迹和RNA过程，使这些酶成为药物开发的潜在关键靶点。然而，由于它们是多结构域蛋白质，并且难以进行结构和功能分析，因此在获得控制其运作模式的结构-功能关系的基本知识方面进展缓慢。为了克服现有的障碍，我们将生产专门的试剂，称为“合成亲和结合剂”（简称sABs），它将用作陪伴物