TOWARDS UNDERSTANDING THE NON-CLASSICAL SECRETION OF HUMAN INTERLEUKIN-1 ALPHA

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 白细胞介素（IL）参与与感染、炎症和自身免疫相关的广泛的生物过程。IL-1特别是促炎细胞因子，它们帮助宿主对抗感染。已知IL-1的缺乏会导致细菌感染的致命性。此外，最近的研究表明，IL-1也在伤口愈合、致癌、肿瘤侵袭、类风湿性关节炎和阿尔茨海默病中发挥作用。IL-1缺乏N-末端信号肽，因此，与大多数其他细胞外蛋白不同，它不通过经典的内质网分泌  高尔基体途径。初步研究表明，在铜的存在下，IL-1 ²通过与钙结合蛋白S100 A13形成多蛋白复合物而释放到细胞外室。虽然存在关于IL-1信号传导过程的有用信息，但IL-1分泌到细胞外区室的确切机制尚不清楚。到目前为止，由于在哺乳动物细胞中过表达IL-1的困难，IL-1的结构-功能数据有限。在这种情况下，拟议研究的目的是充分了解IL-1的非经典释放及其与铜和S100 A13的结合。该提案的具体目标包括：1）确定IL-1 <$对铜和S100 A13的结合亲和力; 2）表征IL-1 <$的非经典分泌所必需的分子相互作用。对IL-1的非经典分泌的完整理解将为理解无信号肽蛋白质输出的一般原理提供有价值的信息。此外，在资助期间，该项目将吸引众多本科生和硕士研究生进行独立研究，提高他们对科学原理的学习，使他们有机会为研究IL-1在这个毁灭性疾病家族中所起的作用做出独特贡献，并激发他们对未来生物医学研究事业的兴奋。此外，该申请的资金将扩大学生在蛋白质稳定性，结构和功能方面的研究，并使更多来自肯塔基州的学生成功进入生物医学研究生课程。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Interleukins (IL's) are involved in a wide spectrum of biological processes associated with infection, inflammation, and autoimmunity. IL-1's in particular are pro-inflammatory cytokines, and they assist the host in fighting infection. Deficiency in IL-1's is known to result in lethality in bacterial infections. In addition, recent studies suggest that IL-1¿ also plays a role in wound healing, carcinogenesis, tumor invasion, rheumatoid arthritis, and Alzheimer's disease. IL-1¿ lacks the N-terminal signal peptide, therefore, unlike most other extracellular proteins, it is not secreted through the classical endoplasmic recticulum  Golgi pathway. Preliminary studies have suggested that the release of IL-1¿ into the extracellular compartment occurs by the formation of a multi-protein complex to the calcium- binding protein, S100A13 in the presence of copper. Although useful information exists on the IL-1 signaling process, the exact mechanism of IL-1¿ secretion into the extracellular compartment is not clear. Until now structure- function data have been limited for IL-1¿ because of the difficulty in its overexpression in mammalian cells. In this context, the purpose of the proposed research is to fully understand IL-1¿'s non-classical release and it's binding to copper and S100A13. Specific aims of this proposal include: 1) determing the binding affinity of IL-1¿ for copper and S100A13; 2) characterizing the molecular interactions necessary for the non-classical secretion of IL-1¿. A complete understanding of the non-classical secretion of IL-1¿ will provide valuable information towards understanding the general principles of export of proteins without signal peptide. Furthermore, over the funding period, this project will engage numerous undergraduates and master's level graduate students in independent research, enhancing their learning of scientific principles, giving them an opportunity to make unique contributions to the study the role played by IL-1¿ in this devastating family of diseases, and stimulating their excitement for future careers in biomedical research. Moreover, the funding of this application will expand student research in protein stability, structure and function and enable more students from Kentucky, a state traditionally underrepresented in biomedical sciences, to successfully advance into biomedical graduate programs.