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）：该项目的总体目标是阐明控制组蛋白修饰酶催化和调节的分子机制。我们将使用支持伴侣的生物学和结构（CEBS）技术平台来研究一个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）及其互补伴侣，赖氨酸脱甲基酶（KDM）通过其作用作为表观遗传信号传导的关键介体，作为其“作家和擦除者”的“作家和擦除者”，对组蛋白上的跨性别型蛋白质的转换后修饰。许多最近的研究强调了组蛋白的赖氨酸甲基化的重要性，导致直接影响DNA复制，修复，重组，基因沉默，印迹和RNA过程，从而使这些酶潜在的药物开发潜在的关键目标。然而，在获得有关其运行方式的结构 - 功能关系的基本知识方面的基本知识方面的进展很慢，因为它们是多域蛋白，并且对结构和功能分析均已顽固。为了克服现有的障碍，我们将生成称为“合成亲和力粘合剂”或将用作伴侣的专业试剂 结晶以及用于细胞生物学应用的定制亲和力试剂。为了实现我们的目标，我们组建了一个世界一流的研究人员团队，该团队将利用SAB试剂来确定结构确定和高水平的生物学测定。我们方法的主要重点是在结构/生化上识别和在结构/生化上表征了分子复合物，其中HMT和KDMS通过使用稳定和增强复合物结晶的SAB来函数。因此，CEBS的努力将依靠与大型高通量中心的紧密联系，以在多种构象状态，酶 - 基底络合物和多蛋白质复合物中的单个酶的蛋白质产生和结构测定。我们方法的一个独特优势是，我们将提供结晶的伴侣和相互作用伙伴的信息到高吞吐量中心，以极大地增加结构确定成功的可能性。 公共卫生相关性：赖氨酸特异性组蛋白甲基转移酶和组蛋白脱甲基酶是DNA复制和修复的表观遗传调节中的关键酶，以及重组以及基因表达和抑制。这些酶的突变与多种疾病有关。这些研究将为药物开发提供生物学原理和结构性基础。