Developing a cell-based high throughput screening for USP15 deubiquitinase inhibitor discovery

USP15脱泛素酶抑制剂的细胞高通量筛选 该项目的目标是开发一种基于泛素探针的AlphaLISA脱泛素酶HTS检测方法， 实现基于细胞的高通量筛选人去泛素酶USP15。现在很明显， 人类泛素系统对细胞和机体的正常功能至关重要。 去泛素酶或DUBS作为一类重要的酶与神经疾病有关， 尤其是帕金森氏症(PD)。许多与疾病相关的潜在可用药去泛素酶是 目前无法获得基于HTS的抑制剂发现，因为难以将其纯化为均相 以及数量足够大的活性重组蛋白，以支持HTS活动。在这项建议中，我们的目标是 用于高通量筛选USP15等的细胞AlphaLISA脱泛素酶检测方法的建立 DUBS，并为鉴定和开发具有促有丝分裂作用的USP15抑制剂奠定了基础 作用和发展成为帕金森病疗法的潜力。

Developing a cell-based high throughput screening for USP15 deubiquitinase inhibitor discovery The goal of this project is to develop a ubiquitin probe-based AlphaLISA deubiquitinase HTS assay that enables cell-based and high throughput screening for human deubiquitinase USP15. It is now clear that the human ubiquitin system is fundamentally important to the normal cellular and organismal functions. Deubiquitinases or DUBs as an important class of enzyme have been associated with neurological disorders, particularly Parkinson’s disease (PD). Many disease-relevant and potentially druggable deubiquitinases are currently inaccessible to HTS-based inhibitor discovery due to the difficulty in their purification as homogeneous and active recombinant proteins in quantity large enough for HTS campaign. In this proposal, we aim at developing a cell-based AlphaLISA deubiquitinase assay for high throughput screening against USP15 and other DUBs, and setting the ground work for identifying and developing USP15 inhibitors with a mitophagy-promoting effect and the potential being developed into PD therapeutics.