Histone Modification Mechanisms and Inhibition

描述(由申请人提供)：本申请是GM62437的竞争性续展申请，涉及化学方法，以筛选修饰组蛋白和其他蛋白质的乙酰基转移酶和去甲基酶的机制、调节和功能。P300/CBP组蛋白乙酰转移酶(HAT)和LSD1组蛋白去甲基酶是调节基因表达的关键酶，是一系列疾病的潜在治疗靶点。P300/CBP是一种广泛作用的HAT，参与调节癌症通路、内分泌紊乱、免疫信号以及学习和记忆。LSD1是一种转录抑制因子，可以从组蛋白H3的Lys4中去除甲基。在这一应用中，我们计划在我们最近对p300/CBP HAT结构域和LSD1催化结构域进行结构分析的基础上，与我们的合成抑制剂形成复合体。具体目标有四个：1)根据p300 HAT的X射线结构确定p300/CBP HAT催化和调控的范围和机制。在这方面，我们将使用突变和表达蛋白连接的方法来研究p300/CBP L1环和自动调节环的作用。2)开发新一代选择性小分子HAT抑制剂，并将其应用于基因调控和疾病模型研究。基于p300/CBP HAT和PCAF/GCN5 HAT在双底物类似物中的晶体结构，将采用虚拟配体筛选。3)确定LSD1组蛋白去甲基酶底物识别的基础。将使用化学和生化策略来辨别底物伽马转向的潜在作用。4)开发用于细胞内LSD1活性调节和用于蛋白质组学分析的合成抑制剂。引线将从丙叉胺和联氨支架开发出来。综上所述，这些研究有可能揭示染色质重塑的基础，并为药物干预确定新的方向。公共卫生相关性：人们对表观遗传学的生物学和药理学越来越感兴趣。这项申请涉及两个关键的酶参与调节的表观遗传染色质重塑。从这项研究计划中获得的见解可能会导致癌症、糖尿病、免疫紊乱和神经精神疾病的新治疗策略。

DESCRIPTION (provided by applicant): This application is a competing renewal application of GM62437 that concerns chemical approaches to sorting out the mechanisms, regulation, and functions of acetyltransferase and demethylase enzymes that modify histones and other proteins. p300/CBP histone acetyltransferase (HAT) and LSD1 histone demethylase are key enzymes that regulate gene expression and are potential therapeutic targets in a range of diseases. p300/CBP is a broadly acting HAT that has been implicated in modulating cancer pathways, endocrine disorders, immune signaling, and learning and memory. LSD1 is a transcriptional repressor that removes methyl groups from Lys4 of histone H3. In this application, we plan to build on our recent progress regarding structural analysis of the p300/CBP HAT domain and the LSD1 catalytic domain in complex with our synthetic inhibitors. There are four specific aims: 1) Determine the scope and mechanisms of p300/CBP HAT catalysis and regulation based on the p300 HAT X-ray structure. In this regard, we will investigate the role of the p300/CBP L1 loop and the autoregulatory loop using mutagensis and by employing expressed protein ligation. 2) Develop a new generation of selective small molecule HAT inhibitors and apply these to gene regulation and disease model studies. Virtual ligand screening will be employed based on the crystal structures of p300/CBP HAT and PCAF/GCN5 HAT in complex with bisubstrate analogs. 3) Determine the basis for LSD1 histone demethylase substrate recognition. Chemical and biochemical strategies will be used to discern the potential role of a substrate gamma turn. 4) Develop synthetic inhibitors for the modulation of LSD1 activity in cells and for use in proteomics analysis. Leads will be developed from propargylamine and hydrazine scaffolds. Taken together, these studies have the potential to uncover important insights into the basis of chromatin remodeling and define new directions for pharmacologic intervention. PUBLIC HEALTH RELEVANCE: There is increasing interest in the biology and pharmacology of epigenetics. This application addresses two key enzymes involved in the regulation of epigenetic chromatin remodeling. Insights obtained from this research plan could lead to new therapeutic strategies for cancer, diabetes, immune disorders, and neuropsychiatric conditions.