AH RECEPTOR SIGNALING MECHANISM

AH 受体信号传导机制

• 批准号: 2810666
• 负责人: WILLIAM K CHAN
• 金额: $ 7.5万
• 依托单位: UNIVERSITY OF THE PACIFIC-STOCKTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2002-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2810666
• 关键词: Baculoviridae; aromatic hydrocarbon receptor; biological signal transduction; dioxins; environmental contamination; gene expression; ligands; protein protein interaction; protein purification; protein structure function; receptor binding; transcription factor; western blottings

DESCRIPTION: (Adapted from the Investigator's Abstract) Dioxins, generated both commercially and naturally, are chlorinated polycyclic aromatic hydrocarbons that are highly toxic environmental contaminants. These agents are known to be potent rodent carcinogens and suspected human carcinogens. The best known prototype of this group of agents is 2,3,7,8-tetrachlorodibenzo-p- dioxin (TCDD). It has been well-documented that most, if not all, of the TCDD effects are mediated through the Ah receptor (AHR). Thus, in an effort to better understand the mechanism of dioxin action, the investigator proposes to investigate the molecular mechanism of the AHR signaling pathway using the overexpressed AHR and ARNT proteins. The investigator's working hypothesis is as follows: Upon ligand binding, the AHR undergoes conformational changes resulting in alteration of a number of AHR-protein interactions, which subsequently leads to a cascade of biologic events to occur. In addition to AHR-protein interactions, other interactions involving ARNT and other proteins also contribute to the action of dioxins mediated by the AHR. This proposal contains specific aims investigating the protein factors and ARNT-associated proteins involved in the AHR signaling pathway. Not only do these studies provide important information and reagents to further the study of the PAH action mediated by the AHR, but they also provide mechanistic insights on how the AHR regulates its target genes by understanding what other proteins are involved in the AHR signaling pathway. Three specific aims have been proposed: Aim 1) purification and functional characterization of baculovirus expressed human AHR; Aim 2) identification and characterization of protein factors required for AHR and ARNT (Ah receptor nuclear translocator) function and Aim 3) investigation of ARNT-associated protein using Far-Western blot analysis.

产品说明：（改编自《研究者摘要》）商业和自然产生的二恶英是氯化多环芳烃，是剧毒的环境污染物。已知这些物质是啮齿类动物的强效致癌物和疑似人类致癌物。这类药剂最著名的原型是2，3，7，8-四氯二苯并对二恶英（TCDD）。它已被充分证明，大多数，如果不是全部，TCDD的影响是通过介导的Ah受体（AHR）。因此，为了更好地理解二恶英的作用机制，研究者建议使用过表达的AHR和ARNT蛋白来研究AHR信号通路的分子机制。研究者的工作假设如下：在配体结合时，AHR经历构象变化，导致许多AHR-蛋白质相互作用的改变，这随后导致发生级联生物事件。除了AHR与蛋白质的相互作用外，ARNT与其他蛋白质的相互作用也有助于AHR介导的二恶英的作用。该计划包含了研究参与AHR信号通路的蛋白质因子和ARNT相关蛋白的具体目标。这些研究不仅为进一步研究AHR介导的PAH作用提供了重要的信息和试剂，而且还通过了解AHR信号通路中涉及的其他蛋白质，为AHR如何调节其靶基因提供了机制见解。提出了三个具体目标：目的1）杆状病毒表达的人AHR的纯化和功能表征;目的2）AHR和ARNT（Ah受体核转运蛋白）功能所需的蛋白质因子的鉴定和表征;目的3）使用Far-Western印迹分析研究ARNT相关蛋白。