Oligomerization State and Function of the NLRP3 Inflammasome

 描述（由申请人提供）：NLRP 3炎性体是哺乳动物先天免疫系统的重要细胞内组分。在过去的十年中，重要的研究已经极大地提高了炎症体如何对大量不同的细菌，病毒，真菌和内源性病原体做出反应的知识。炎性小体激活的关键步骤是三种蛋白质的复合物形成和寡聚化。然而，确切的寡聚化状态和这种寡聚化状态与炎性小体功能的关系尚未确定。本申请中描述的拟议研究将在哺乳动物细胞系统中产生三种炎性体蛋白，并探索这些问题。通过研究驱动NLRP 3炎性小体激活的结构机制，我们希望进一步全面了解先天免疫系统的这一关键组成部分，并确定可能用于治疗调节的关键因素。关于这些机制的新信息将对神经变性、心脏病和许多炎症性疾病的治疗产生影响。 本申请中描述的拟议研究将在哺乳动物细胞系统中表达三种重组炎性体蛋白，并研究每种蛋白的寡聚化状态。此外，每个蛋白质伴侣和整个三元复合物之间的相互作用将被定义，并确定其组织。最后，我们将阐明高阶寡聚化状态如何影响炎性小体功能。拟议的研究将是知识的重要一步，将指导未来对这一重要炎症调节因子的研究，并将为许多对生物化学，生物物理和生物医学研究感兴趣的学生提供巨大的教育参与和推广。

 DESCRIPTION (provided by applicant): The NLRP3 inflammasome is an important intracellular component of the mammalian innate immune system. Significant research over the last decade has vastly improved the knowledge of how the inflammasome responds to a large and diverse group of bacterial, viral, fungal, and endogenous pathogens. A key step in inflammasome activation is complex formation and oligomerization of three proteins. However, the exact oligomerization state and the relationship of this oligomerization state to inflammasome function have not yet been determined. The proposed research study described in this application will produce the three inflammasome proteins in a mammalian cell system and probe these questions. By investigating the structural mechanisms driving NLRP3 inflammasome activation, we hope to further the overall understanding of this critical component of the innate immune system and identify key elements that might be exploited for therapeutic modulation. New information on these mechanisms will have implications for the treatment of neurodegeneration, heart disease, and numerous inflammatory diseases. The proposed research study described in this application will express three recombinant inflammasome proteins in a mammalian cell system and investigate the oligomerization state of each protein. Furthermore, interactions between each protein partner and the whole ternary complex will be defined and their organization determined. Finally, we will elucidate how the higher-order oligomerization state impacts inflammasome function. The proposed studies will be a significant step forward in knowledge, will guide future investigations of this important inflammatory regulator, and will provide tremendous educational involvement and outreach for many students interested in biochemical, biophysical, and biomedical studies.