Protein Acylation and Methylation Mechanisms

蛋白质酰化和甲基化机制

• 批准号: 10361462
• 负责人: PHILIP A COLE
• 金额: $ 42.96万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10361462
• 关键词: Acetylation; Acylation; Acyltransferase; Alkylation; Area; Binding; Biochemical; Biological Assay; Biological Process; Cell Culture Techniques; Cell physiology; Cellular Assay; Chemicals; Coenzyme A; Development; Diabetes Mellitus; Disease; Drug Targeting; EP300 gene; Enzyme Inhibitor Drugs; Enzymes; Epigenetic Process; Epitopes; Excision; Feedback; Gene Expression Regulation; Histone H3; Histones; Homocysteine; Hormones; Hydrolase; In Vitro; Inositol; Investigation; KDM1A gene; Knowledge; Lead; Ligation; Lipids; Lysine; Malignant Neoplasms; Maps; Membrane; Metabolic; Metabolic Diseases; Metabolism; Methods; Methylation; Methyltransferase; Modification; Molecular; Nucleotides; Oxidoreductase; PCAF gene; Pathogenesis; Peptides; Phenelzine; Positioning Attribute; Post-Translational Protein Processing; Post-Translational Regulation; Process; Protein Acetylation; Protein Engineering; Protein Methyltransferases; Protein Microchips; Proteins; Reaction; Regulation; Reporting; Research; Role; S-Adenosylmethionine; Shapes; Side; Signal Transduction; Site; Structure; Substrate Interaction; Techniques; Testing; analog; cancer therapy; chromatin remodeling; crosslink; demethylation; diagnostic strategy; disease diagnostic; enzyme substrate complex; experimental study; ghrelin; histone acetyltransferase; histone demethylase; improved; in vivo; inhibitor; methyl group; molecular recognition; neoplastic; novel therapeutic intervention; novel therapeutics; obesity treatment; peptide hormone; small molecule; therapeutic target; tool

This proposal is a competing renewal application of R01 GM62437 that combines the use of chemical approaches, enzymologic analysis, and cellular studies to enhance our understanding of enzymes regulating protein acylation and methylation. It is now well-accepted that post-translational modifications (PTMs) involving lysine acetylation and reversible methylation on histones and other proteins are central to epigenetics. Such epigenetic modifying enzymes are viewed as attractive drug targets for cancer and other diseases. The ghrelin O-acyltransferase (GOAT) enzyme catalyzes the unusual PTM of octanoyl attachment to a Ser side chain of the peptide hormone ghrelin and inhibitors of this enzyme may be useful in the treatment of obesity and diabetes. Although there has been increasing efforts to understand the mechanisms and functions of these PTMs and the enzymes that catalyze them, there are major gaps in our understanding in these areas. Filling these knowledge gaps has the potential to provide a clearer understanding of basic biomedical processes and has the opportunity to enhance the development of novel therapeutic approaches and disease diagnostic strategies. There are four Specific Aims in this proposal. 1. Elucidate the molecular basis for protein substrate selectivity for histone acetyltransferases (HAT) using new chemical and biochemical approaches. We will develop and apply new techniques to generate histone-CoA conjugates and exploit protein microarrays to clarify structural and functional features of HAT-substrate interactions. 2. Develop and apply improved chemical tools for analyzing LSD1 histone demethylation. A combination of synthetic inhibitors and propargyl-histones will be prepared to interrogate LSD1 cellular functions and molecular interactions. 3. Clarify the effects of Lys acetylation on S-adenosyl homocysteine hydrolase and inositol monophosphate dehydrogenase-2. Expressed protein ligation will be used to install acetyl-Lys site-specifically into these metabolic enzymes to dissect potentially important nodes between protein acetylation and cellular metabolism. 4. Define major structural features of ghrelin O-acyltransferase. A combination of membrane topology mapping and bisubstrate analog crosslinking will be used to create a blueprint of this key metabolic regulatory enzyme. We believe that this research effort has the potential to greatly expand our understanding of protein post-translational modification mechanisms and functions and identify new therapeutic opportunities for treating metabolic and neoplastic diseases.

该提案是 R01 GM62437 的竞争性更新申请，结合了使用 化学方法、酶学分析和细胞研究，以增强我们的 了解调节蛋白质酰化和甲基化的酶。现在已被广泛接受 涉及赖氨酸乙酰化的翻译后修饰（PTM）和 组蛋白和其他蛋白质上的可逆甲基化是表观遗传学的核心。这样的 表观遗传修饰酶被视为治疗癌症和其他疾病的有吸引力的药物靶点 疾病。生长素释放肽 O-酰基转移酶 (GOAT) 酶催化以下不寻常的 PTM 辛酰基连接到肽激素 ghrelin 的 Ser 侧链和抑制剂 这种酶可能有助于治疗肥胖和糖尿病。虽然有 正在加紧努力了解这些 PTM 的机制和功能， 对于催化它们的酶，我们在这些领域的理解存在重大差距。 填补这些知识空白有可能使人们更清楚地了解基本知识 生物医学过程，并有机会加强新型药物的开发 治疗方法和疾病诊断策略。有四个具体目标 这个建议。 1. 阐明蛋白质底物选择性的分子基础 使用新的化学和生化方法的组蛋白乙酰转移酶（HAT）。我们 将开发和应用新技术来生成组蛋白-CoA 缀合物并利用 蛋白质微阵列阐明 HAT 底物的结构和功能特征 互动。 2. 开发并应用改进的化学工具来分析LSD1组蛋白 去甲基化。合成抑制剂和炔丙基组蛋白的组合将 准备探究 LSD1 细胞功能和分子相互作用。 3. 澄清 赖氨酸乙酰化对S-腺苷同型半胱氨酸水解酶和肌醇的影响 单磷酸脱氢酶-2。表达的蛋白连接将用于安装 乙酰基赖氨酸位点特异性地进入这些代谢酶以剖析潜在的重要作用 蛋白质乙酰化和细胞代谢之间的节点。 4. 定义主要结构 生长素释放肽O-酰基转移酶的特点。膜拓扑映射的组合 双底物模拟交联将用于创建该密钥的蓝图 代谢调节酶。我们相信这项研究工作有潜力 极大地扩展了我们对蛋白质翻译后修饰机制的理解 和功能，并确定治疗代谢和疾病的新治疗机会 肿瘤性疾病。

项目成果

Regulation of Myf5 Early Enhancer by Histone Acetyltransferase p300 during Stem Cell Differentiation.

干细胞分化过程中组蛋白乙酰转移酶 p300 对 Myf5 早期增强子的调节。

• DOI: 10.4172/2168-9547.1000103
• 发表时间: 2012
• 期刊: Molecular biology
• 影响因子: 1.2
• 作者: Francetic,Tanja;LeMay,Melanie;Hamed,Munerah;Mach,Hymn;Meyers,David;Cole,PhilipA;Chen,Jihong;Li,Qiao
• 通讯作者: Li,Qiao

Site-Specific 5-Formyl Cytosine Mediated DNA-Histone Cross-Links: Synthesis and Polymerase Bypass by Human DNA Polymerase η.

• DOI: 10.1002/anie.202109418
• 发表时间: 2021-12-13
• 期刊: Angewandte Chemie (International ed. in English)
• 作者: Pujari SS;Wu M;Thomforde J;Wang ZA;Chao C;Olson NM;Erber L;Pomerantz WCK;Cole P;Tretyakova NY
• 通讯作者: Tretyakova NY

A selective phenelzine analogue inhibitor of histone demethylase LSD1.

• DOI: 10.1021/cb500018s
• 发表时间: 2014-06-20
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Prusevich, Polina;Kalin, Jay H.;Ming, Shonoi A.;Basso, Manuela;Givens, Jeffrey;Li, Xin;Hu, Jianfei;Taylor, Martin S.;Cieniewicz, Anne M.;Hsiao, Po-Yuan;Huang, Rong;Roberson, Heather;Adejola, Nkosi;Avery, Lindsay B.;Casero, Robert A., Jr.;Taverna, Sean D.;Qian, Jiang;Tackett, Alan J.;Ratan, Rajiv R.;McDonald, Oliver G.;Feinberg, Andrew P.;Cole, Philip A.
• 通讯作者: Cole, Philip A.

Histone H2B Deacylation Selectivity: Exploring Chromatin's Dark Matter with an Engineered Sortase.

• DOI: 10.1021/jacs.1c13555
• 发表时间: 2022-03-02
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Wang ZA;Whedon SD;Wu M;Wang S;Brown EA;Anmangandla A;Regan L;Lee K;Du J;Hong JY;Fairall L;Kay T;Lin H;Zhao Y;Schwabe JWR;Cole PA
• 通讯作者: Cole PA

Histone deacetylase inhibitors decrease NHEJ both by acetylation of repair factors and trapping of PARP1 at DNA double-strand breaks in chromatin.

• DOI: 10.1016/j.leukres.2016.03.007
• 发表时间: 2016-06
• 期刊: LEUKEMIA RESEARCH
• 影响因子: 2.7
• 作者: Robert, Carine;Nagaria, Pratik K.;Pawar, Nisha;Adewuyi, Adeoluwa;Gojo, Ivana;Meyers, David J.;Cole, Philip A.;Rassool, Feyruz V.
• 通讯作者: Rassool, Feyruz V.