Investigation of the Functional/Folding Landscapes of the Il-1 Family

描述（由申请人提供）：先前拨款的主要目的是使用生物物理和计算技术的组合来评估17 KD β蛋白白介素-1 β (il -1 β)折叠/错误折叠的关键特征。我们已经确定，这种β -三叶草蛋白的缓慢折叠在很大程度上是由“拓扑挫折”控制的。在这段时间里，我们也对理解构象动力学在这个蛋白质家族的生物学功能中所起的作用产生了兴趣。从“研究β蛋白的折叠机制”到“研究IL-1家族的功能/折叠景观”的标题变更反映了这一研究重点的变化。此外，我们还增加了何塞·奥努奇奇博士作为这项建议的共同负责人。我们对我们希望解决的问题确立了一种综合的实验/理论方法。很明显，在细菌感染和脓毒性休克中，il -1 β加工的调控是至关重要的；自身免疫性疾病和某些癌症1-8。然而，对il -1 β的加工和成熟的分子要求知之甚少。本提案中概述的研究目标是：(a)研究该蛋白家族中功能/折叠景观的相互作用(b)了解31 KD前体蛋白切割的结构基础；(c) 31 KD前体蛋白和24 KD部分裂解蛋白的结构特征；(d)了解这些序列在破坏成熟的17kd蛋白折叠中的作用。总之，我们的目标是研究相关蛋白的功能格局，通过分析细胞因子激动剂和拮抗剂受体复合物的能量格局，开发理解受体结合和信号传递的方法，并探索proll -1 β从非活性部分结构化状态到活性成熟的il -1 β过程中的构象变化。这些研究在理解蛋白质识别和活性方面具有重要意义，在理解调节prol -1 - β活化的构象过程方面也具有重要意义。

DESCRIPTION (provided by applicant): The principal aim of the previous grant was to use a combination of biophysical and computational techniques to evaluate the key features in the folding/misfolding of the 17 KD beta protein interleukin-1beta (IL-1beta). We have established that the slow folding of this beta-trefoil protein is largely controlled by "topological frustration". During this time period we also became interested in understanding the role conformational dynamics plays in the biological function of this family of proteins. The change in the Title of the grant from "Investigation of the Folding Mechanism of a Beta Protein" to "Investigation of the Functional/Folding Landscapes of the IL-1 Family" reflect this change in research focus. In addition, we have added Dr. Jose Onuchic as a co-Pi on this proposal we have established an integrated experimental/theoretical approach towards the questions we wish to address. It is apparent that the regulation of the processing of IL-1beta is of central importance in bacterial infection and septic shock; autoimmune diseases and some cancers1-8. Yet, little is known about the molecular requirements for processing and maturation of IL-1beta. The goals of the research outlined in this proposal are directed towards (a) investigating the interplay of the functional/folding landscapes within this family of proteins (b) understanding the structural basis for cleavage of the 31 KD precursor protein; (c) the structural characterization of the 31 KD precursor protein and the 24 KD partially cleaved protein; (d) understanding the role of the presequences in destabilizing the fold of the mature 17 KD protein. In summary, our goals are to look at the functional landscapes for related proteins, to develop methods for understanding the receptor binding and signal transmission by analyzing the energy landscapes for the cytokine agonist and antagonist receptor complexes, and exploring the conformational changes that accompany the processing of prolL-1beta from the inactive partially structured state to the active, mature IL-1beta. These studies are of both fundamental importance in understanding protein recognition and activity as well as of fundamental biological importance in understanding the conformational processes that regulate the activation of prolL-1beta.