DESIGN OF MULTIMERIC INTERLEUKIN-2 RECEPTOR ECTODOMAINS

描述：白细胞介素2是一种广泛存在的螺旋细胞因子。 某些恶性肿瘤和感染性疾病的临床应用 包括艾滋病毒感染在内的疾病。白介素2受体(IL-2R) 造血素大家族中最复杂的受体系统 感受器。它可能存在于各种亲和力状态中，具体取决于 三个独立细胞表面的不同组合的参与 亚基：IL-2Rα、β和γ。这些亚单位在形式中起作用 两个特定的杂二聚体(IL-2Rαβ)和(IL-2Rβ伽马)和一个 杂三聚体(IL-2Rαβ-γ)。这些细胞表面的每一个组件 具有独特的生理功能。正是这些因素的协作性 在配体捕获和信号传输过程中提供关键的亚基 以了解开发IL-2的治疗潜力。这个项目 是第一个提出并实施使用卷绕线圈识别的人 复杂受体溶液组装的序列(亮氨酸拉链) 系统。在最初的项目阶段，这一点的可行性 建立了方法，发展了表达和表达的方法 可溶性IL-2R螺旋卷曲复合体的表征。盘绕线圈 介导性溶液组装产生结合的异构体IL-2R复合体 配基的方式类似于细胞表面的对应物。这些 络合物已被证明对IL-2结构/功能分析有用。 本提案是对继续这项工作的请求，目标是 稳定的解决方案组装和完整的表征所有三个 功能异构体IL-2R复合体。这个项目的具体目标是 主要内容如下：1.表达、分离和鉴定两种功能的IL-2 受体异二聚体(IL-2Rβ假性高亲和力和IL-2Rβ 伽马中间亲和力受体)作为稳定的螺旋线圈复合体。2. 重组人IL-2受体的表达、分离和鉴定 异三聚体高亲和力受体(IL-2Rαβ)作为一种稳定的 盘绕线圈复合体。3.测定平衡配基和动力学配基 每个受体复合体的结合特性。4.获取详细信息 通过与这些配合物共结晶得到它们的结构-活性数据 IL-2，并确定其高分辨率的X射线结构。

DESCRIPTION: Interleukin-2 is a helical cytokine that has found wide spread clinical use in the treatment of some malignancies and for infectious diseases including HIV infection. The interleukin-2 receptor (IL-2R) is one of the most complex receptor systems of the large family of hematopoietin receptors. It may exist in a variety of affinity states depending upon the participation of different combinations of three separate cell surface subunits: IL-2Ralpha, beta and gamma. These subunits function in the form of two specific heterodimers (IL-2Ralpha beta) and (IL-2Rbeta gamma) and one heterotrimer (IL-2Ralpha beta gamma). Each of these cell surface assemblies performs a distinct physiologic function. It is the cooperativity of these subunits during ligand capture and signal transmission that provides the key to understanding exploiting the therapeutic potential of IL-2. This project was the first to propose and implement the use of coiled-coil recognition sequences (Leu Zippers) for the solution assembly of complex receptor systems. During the initial project period, the feasibility of this approach was established and methods were developed for the expression and characterization of soluble IL-2R coiled-coil complexes. Coiled-coil mediated solution assembly generated heteromeric IL-2R complexes that bound ligand in a fashion that resembled their cell surface counterparts. These complexes are already proving useful for IL-2 structure/function analysis. This proposal is a request for continuation of this work with the goals of stable solution assembly and complete characterization of all three of the functional heteromeric IL-2R complexes. The Specific Aims of this project are: 1. To express, isolate and characterize the two functional IL-2 receptor heterodimers (IL-2Ralpha beta pseudo high affinity and IL-2R beta gamma intermediate affinity receptors) as stable coiled-coil complexes. 2. To express, isolate and characterize the presumed IL-2 receptor heterotrimeric high affinity receptor (IL-2Ralpha beta gamma) as a stable coiled-coil complex. 3. To determine the equilibrium and kinetic ligand binding characteristics of each receptor complex. 4. To obtain detailed structure-activity data on these complexes by co-crystallizing them with IL-2 and determining their X-ray structures to high resolution.