Deubiquitinating enzymes (DUBs) are a largely understudied and untapped resource in the toolkit of protein degradation functionalities. They comprise a large repertoire of enzymes that remove ubiquitin from substrates in a variety of cellular and pathophysiological contexts, and have enormous potential for research and clinical use. It is only within the last 5 years that potent, selective, and well-characterized small-molecule inhibitors of DUBs have been described. These compounds are now being used to study the biological roles of DUBs. Here, we describe downstream applications of small-molecule inhibitors for studying DUBs and provide a framework for future studies. We highlight recent examples of using these inhibitors to confirm and explore the role of these enzymes in both normal and pathological contexts. These studies represent the first steps in the burgeoning field of pharmacological and chemoproteomic studies of DUBs, which will be critical for the continued advancement of DUB field.

去泛素化酶 (DUB) 是蛋白质降解功能工具包中的一种很大程度上尚未得到研究和开发的资源。它们包含大量酶，可在各种细胞和病理生理背景下从底物中去除泛素，具有巨大的研究和临床应用潜力。仅在最近 5 年内才描述了 DUB 的有效、选择性和充分表征的小分子抑制剂。这些化合物现在被用于研究 DUB 的生物学作用。在这里，我们描述了小分子抑制剂在研究 DUB 中的下游应用，并为未来的研究提供了一个框架。我们重点介绍了最近使用这些抑制剂来确认和探索这些酶在正常和病理环境中的作用的例子。