UNC Center for Nanosensing the Stressome (RMI)

作为响应细胞压力的一个单位，蛋白激酶的支架组装构成了一种重要的纳米机器，称为应激体。应激体的适当功能对体内平衡至关重要，在包括炎症和缺血在内的许多人类疾病中都处于失调状态。压力不是简单的信号相互作用的集合，而是一个实际的大型多蛋白质复合体，其不断变化的组成和亚细胞位置决定功能。对应激的有效反应还需要对应激体内蛋白质的激活和失活进行精确的时空协调。要监视 动态变化需要创建识别、报告并可能阻断应激信号通路的激活蛋白的探针。因此，我们正在开发不仅识别特定蛋白质而且区分这些蛋白质的不同构象状态的探针。这些探针目前是小的蛋白质结构域，我们称之为构象特异性识别元件。最近的一项突破是能够将构象特定的识别元件共价连接到溶剂敏感的 染料是已知的最亮的染料之一，极大地增强了检测非常低丰度的内源信号分子的灵敏度。除了改进小蛋白结构域的用途外，我们还建议使用组合方法来分离小肽和合成的小分子构象特异性识别元件来检测应激蛋白的活性和非活性构象。蛋白质构象传感探针被连接到新的细胞穿透性多肽上，以传递到活细胞以成像信号转导纳米机器。生物学家、化学家、数学家和物理学家正在生物传感器、蛋白质设计、多肽和小分子发现等领域共同努力， 纳米设备的开发和交付、计算建模、疾病的动物模型和 计划/监督在纳米医学领域创建北卡罗来纳大学的倡议。关于人类疾病，我们的主要长期目标将是在缺血性心血管疾病领域，但我们的工作将影响许多疾病的诊断和治疗。最后，北卡罗来纳大学纳米医学中心将积极与其他纳米医学中心和著名的生物医学研究人员互动，研究生物纳米机器及其在疾病和治疗反应中的作用。

The scaffolded assemblies of protein kinases that function as a unit in responding to cellular stress constitute an important nanomachine termed the stressome. Proper function of the stressome is critical for homeostasis and is dysregulated in many human diseases including inflammation and ischemia. The stressome is not simply a collection of signaling interactions, but an actual large multi-protein complex, whose changing composition and subcellular location dictates function. Efficient response to stress also requires precise spatial and temporal coordination for activation and inactivation of proteins within the stressome. To monitor dynamic changes requires the creation of probes that recognize, report on, and possibly interdict activated proteins of stress signaling pathways. Thus, we are developing probes that not only recognize specific proteins but that discriminate between different conformational states of those proteins. These probes are currently small protein domains that we refer to as conformation specific recognition elements. A recent breakthrough has been the ability to covalently attach conformation specific recognition elements to solvent-sensitive dyes that are among the brightest dyes known, providing greatly enhanced sensitivity to examine very low abundance, endogenous signaling molecules. In addition to refining the use of small protein domains we propose using combinatorial methods to isolate small peptide and synthetic small molecule conformation-specific recognition elements for detecting active and inactive conformations of stress proteins. Protein conformation sensing probes are coupled to novel cell-penetrating peptides for delivery to living cells to image signal transduction nanomachines. Biologists, chemists, mathematicians and physical scientists are working together in the areas of biosensors, protein design, peptide and small molecule discovery, nanodevice development and delivery, computational modeling, animal models of disease and planning/oversight to create the UNC initiative in nanomedicine. Relative to human disease our primary long term goal will be in the area of ischemic cardiovascular disease but our work will impact diagnosis and treatment of many diseases. Finally, the UNC Nanomedicine Center will actively interact with other nanomedicine centers and renowned biomedical investigators for the study of biological nanomachines and their role in disease and response to therapy.