The discovery of small molecules to dissect the individual roles of activator-coactivator complexes

发现小分子来剖析激活剂-共激活剂复合物的各个作用

• 批准号: 8987175
• 负责人: Jean Marie Lodge
• 金额: $ 3.5万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8987175
• 关键词: Activator Appliances; Alzheimer' s Disease; Binding; Binding Sites; Biological Process; CREB1 gene; Cells; Chemicals; Complex; Coupled; Cysteine; Disease; Disulfides; Docking; EP300 gene; Event; Fluorescence Polarization; Gene Expression; Genes; Hematopoiesis; Homeostasis; Homologous Gene; Huntington Disease; In Vitro; Individual; Ligands; Link; Lipids; MYB gene; Metabolism; Modeling; Neuronal Plasticity; Physiological; Play; Positioning Attribute; Proteins; Proto-Oncogene Proteins c-myb; Regulation; Research; Role; SREBP-1a; Site; Specificity; Surface; Synaptic plasticity; Therapeutic; Transcription Coactivator; Transcriptional Activation; Western Blotting; Work; analog; cell growth; design; disulfide bond; inhibitor/antagonist; interest; leukemia; liquid chromatography mass spectrometry; mutant; novel; public health relevance; research study; small molecule; tool

 DESCRIPTION (provided by applicant): In the activation of transcription the GACKIX motif, a small conserved domain plays an important role with two potential docking sites for activators and has been found in CBP and its homolog p300 as well as the unrelated coactivator ARC105. The functions of activator*GACKIX domain complexes are diverse and include cellular growth, hematopoiesis, synaptic plasticity, and lipid homeostasis; thus, dysregulation of these complexes are also linked to many diseases from leukemia to Alzheimer's and Huntington's disease. Chemical tools are sought to dissect the individual functions of activator*GACKIX domain complexes and to provide a potential therapeutic model. However small molecules have yet to be designed that can distinguish between the GACKIX domain of a particular coativator such as p300 versus CBP. In this research plan the site-directed strategy known as Tethering is applied to match orthogonal disulfide-containing fragments with cysteine-modified GACKIX (cysGACKIX) domain. First a liquid chromatography-mass spectrometry Tethering screen was used to identify disulfide fragments that target the MLL-binding site of the cysGACKIX domain. A novel fluorescence polarization Tethering screen streamlined the identification of small molecules inhibitors directed at the c-Myb and CREB- binding site of the cysGACKIX domain. The disulfide fragments will be converted into covalent irreversible probes. Tethering will be extended using the most selective and potent small molecules inhibitors to assess the function of activators such as CREB, and MLL interactions with GACKIX domain of CBP, p300, and ARC105 in cells.

 描述（由适用提供）：在转录激活Gackix基序中，一个小的配置域在两个潜在的扩展位点起着激活剂的潜在对接位点的重要作用，并且在CBP及其同源p300以及无关的共激活器ARC105中已发现。激活剂*Gackix结构域复合物的功能是潜水员，包括细胞生长，造血，突触可塑性和脂质稳态；因此，这些复合物的失调也与从白血病到阿尔茨海默氏病和亨廷顿氏病的许多疾病有关。感觉到化学工具以剖析激活剂*Gackix域复合物的各个功能并提供潜在的治疗模型。但是，尚未设计小的分子，可以区分特定盖子的Gackix结构域，例如P300与CBP。在本研究计划中，将被称为绑定的站点定向的策略应用于含二硫二硫键的片段与半胱氨酸修饰的Gackix（Cysgackix）域匹配。首先，使用液相色谱 - 质谱束缚筛网来识别针对Cysgackix域的MLL结合位点的二硫键片段。一个新型的荧光偏振筛网简化了针对Cysgackix结构域C-Myb和Creb结合位点的小分子抑制剂的鉴定。二硫化物片段将转换为共价性不可逆问题。将使用最选择性和潜在的小分子抑制剂扩展束缚，以评估CREB等激活剂的功能，并与CBP，P300和ARC105的Gackix结构域相互作用。