Proteomics and the dioxin signaling network

蛋白质组学和二恶英信号网络

• 批准号: 6657397
• 负责人: JOHN J LAPRES
• 金额: $ 22.43万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-10 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6657397
• 关键词: SDS polyacrylamide gel electrophoresis; aromatic hydrocarbon receptor; biological signal transduction; cytoplasm; dioxins; environmental exposure; immunoprecipitation; mass spectrometry; mitogen activated protein kinase; phosphorylation; physical chemical interaction; polymerase chain reaction; posttranslational modifications; protein protein interaction; proteomics; tissue /cell culture; transfection; western blottings

DESCRIPTION (provided by applicant) 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is characterized as one of the most toxic chemicals known to man. Its toxic effects include porphyria, a severe wasting syndrome, metaplasia, teratogenesis and immune suppression. The aryl hydrocarbon receptor (AHR) is essential to this toxicity. In the absence of ligand, the AHR is found in the cytosol bound to a dimmer of Hsp90 and a monomer of ARAS. ARA9 shares sequence similarities to FK506 binding proteins. ARA9 acts to maintain stability and proper cellular compartmentalization of the AHR. The method by which it fulfills these functions is unknown. In addition, what role ARA9 plays in secondary signaling and toxicity of ligands for the AHR has not been determined. Finally, it is not known whether ARA9 can recruit other cellular proteins to the AHR cytosolic complex or act to influence other cellular signaling cascades. Preliminary data suggests that ARA9 may interact with a known mediator of various signaling pathways. This interaction has lead to the following hypothesis: ARA9 influences dioxin mediated signaling by recruiting other proteins to the AHR cytosolic complex and directly influencing other cellular signaling cascades. To address this hypothesis, three specific aims are proposed: 1) Map the interaction surfaces of ARA9 and other cellular signaling molecules. 2) Use proteomic tools to identify new members of the AHR signaling network. 3) Identify changes in phosphorylation patterns in TCDD treated cells using a novel phosphoproteomic solid phase enrichment technology. Upon completion of these aims, the investigators will have extended the understanding of TCDD toxicity by identifying other cellular targets for signaling. In addition, they will have directly related a significant post-translational modification to this signaling cascade. Finally, this work will establish a potential mechanism for secondary toxicity as it relates to TCDD exposure.

描述（由申请人提供） 2,3,7,8-四氯二苯并-对二恶英 (TCDD) 是人类已知的毒性最强的化学物质之一。其毒性作用包括卟啉症、严重消耗综合征、化生、致畸和免疫抑制。芳烃受体（AHR）对于这种毒性至关重要。在没有配体的情况下，AHR 存在于与 Hsp90 二聚体和 ARAS 单体结合的胞质溶胶中。 ARA9 与 FK506 结合蛋白具有序列相似性。 ARA9 的作用是维持 AHR 的稳定性和适当的细胞区室化。它实现这些功能的方法尚不清楚。此外，ARA9 在 AHR 配体的次级信号传导和毒性中所起的作用尚未确定。 最后，尚不清楚 ARA9 是否可以招募其他细胞蛋白至 AHR 胞质复合物或影响其他细胞信号级联。 初步数据表明，ARA9 可能与多种信号通路的已知介质相互作用。这种相互作用导致了以下假设：ARA9 通过将其他蛋白质招募到 AHR 胞质复合物并直接影响其他细胞信号级联反应来影响二恶英介导的信号传导。为了解决这一假设，提出了三个具体目标：1）绘制 ARA9 和其他细胞信号分子的相互作用表面。 2) 使用蛋白质组学工具识别 AHR 信号网络的新成员。 3) 使用新型磷酸蛋白质组固相富集技术识别 TCDD 处理细胞中磷酸化模式的变化。 完成这些目标后，研究人员将通过识别其他细胞信号传导靶标来扩展对 TCDD 毒性的理解。此外，他们还将显着的翻译后修饰与该信号级联直接相关。 最后，这项工作将建立与 TCDD 暴露相关的二次毒性的潜在机制。