RP5: Chemical proteomic discovery of small-molecule probes for autophagy proteins

项目总结--报告5 增强自噬的小分子有可能用作治疗系统发育疾病的药物 病原体种类繁多，种类繁多。尽管如此，大多数参与或调节的蛋白质 自噬缺乏化学探针，有些甚至被认为是无法下药的。为了加速发现 化学探针，以及最终促进抗感染自噬的药物，需要新的平台来 可以广泛和有效地评估来自不同物种的自噬相关蛋白的小分子相互作用 结构类和功能类。我们实验室最近推出了第一个化学蛋白质组 直接在本地生物系统中全球评估蛋白质的可药性的平台。这些 定量质谱仪-基于活性的蛋白质图谱(MS-ABPP)平台可应用于任何细胞 类型或状态，并在广泛的范围内识别了数百种蛋白质的小分子HIT配体 包括那些以前被认为不能用药的类别(例如，接头、转录因子)。挖掘我们的 目前人类蛋白质组的MS-ABPP数据集包含10,000多个蛋白质上的30,000个+位点，具有 发现了几种与降解自噬和/或ATG基因有关的蛋白质的可药性证据- 依赖豁免权。在Rp5中，我们将与RP1-4密切合作，使用我们的MS-ABPP平台来发现 优化小分子探针，增强自噬介导的对传染病的防御。 我们将追求以下具体目标：1)蛋白质靶标的化学蛋白质组学鉴定 自噬刺激小分子；2)HIT配体的化学蛋白质组学发现 自噬蛋白；3)化学蛋白质组学发现调节自噬的新的可用药蛋白； 4)优先自噬蛋白化学探针的优化。RP5应该提供强有力的和 几种优先自噬蛋白的选择性化学探针，这些探针将被提供给RP1- 4用于生物学研究。在Rp5中开发的化学探针也应该是非常必要的工具 更大的自噬和传染病社区，以及 开发广谱抗感染疗法。

PROJECT SUMMARY – RP5 Small molecules that enhance autophagy have the potential to serve as therapeutics against a phylogenetically diverse and broad range of pathogens. Nonetheless, most of the proteins that participate in, or regulate, autophagy lack chemical probes and some may even be viewed as undruggable. To accelerate the discovery of chemical probes, and ultimately drugs that promote anti-infective autophagy, new platforms are needed that can broadly and efficiently evaluate small molecule interactions for autophagy-related proteins from diverse structural and functional classes. Our laboratory has recently introduced the first chemical proteomic platforms to globally assess the druggability of proteins directly in native biological systems. These quantitative mass spectrometry-activity-based protein profiling (MS-ABPP) platforms can be applied to any cell type or state and have identified small-molecule hit ligands for many hundreds of proteins across a wide range of classes, including those previously considered undruggable (e.g., adaptors, transcription factors). Mining our current MS-ABPP data sets of the human proteome, which contain 30,000+ sites on 10,000+ proteins, has uncovered evidence of druggability for several proteins involved in degradative autophagy and/or ATG gene- dependent immunity. In RP5, we will use our MS-ABPP platforms in close collaboration with RP1-4 to discover and optimize small-molecule probes that enhance autophagy-mediated defense against infectious diseases. We will pursue the following Specific Aims: 1) the chemical proteomic identification of protein targets of autophagy-stimulating small molecules; 2) the chemical proteomic discovery of hit ligands for prioritized autophagy proteins; 3) the chemical proteomic discovery of novel, druggable proteins that regulate autophagy; and 4) the optimization of chemical probes for prioritized autophagy proteins. RP5 should furnish potent and selective chemical probes for several prioritized autophagy proteins, and these probes will be provided to RP1- 4 for biological studies. The chemical probes developed in RP5 should also serve as much needed tools for the greater autophagy and infectious disease communities, as well as valuable starting points for the development of broad-spectrum anti-infective therapies.