In recent years, phenotypic-based screens have become increasingly popular in drug discovery. A major challenge of this approach is that it does not provide information about the mechanism of action of the hits. This has led to the development of multiple strategies for target deconvolution. Thermal proteome profiling (TPP) allows for an unbiased search of drug targets and can be applied in living cells without requiring compound labeling. TPP is based on the principle that proteins become more resistant to heat-induced unfolding when complexed with a ligand, e.g., the hit compound from a phenotypic screen. The melting proteome is also sensitive to other intracellular events, such as levels of metabolites, post-translational modifications and protein-protein interactions. In this review, we describe the principles of this approach, review the method and its developments, and discuss its current and future applications. While proteomics has generally focused on measuring relative protein concentrations, TPP provides a novel approach to gather complementary information on protein stability not present in expression datasets. Therefore, this strategy has great potential not only for drug discovery, but also for answering fundamental biological questions.

中文翻译：

---

热蛋白质组分析：通过热诱导的稳定性变化无偏评估蛋白质状态。

近年来，基于表型的筛选在药物发现中越来越受欢迎。这种方法的主要挑战是它不提供有关命中的作用机理的信息。这导致了针对目标反卷积的多种策略的发展。热蛋白质组分析（TPP）可以无偏倚地搜索药物靶标，并且可以在活细胞中使用而无需化合物标记。TPP基于以下原理：蛋白质与配体（例如，来自表型筛选的命中化合物）复合时，对热诱导的展开更具抵抗力。融化的蛋白质组还对其他细胞内事件敏感，例如代谢物的水平，翻译后修饰和蛋白质-蛋白质相互作用。在这篇评论中，我们描述了这种方法的原理，回顾该方法及其发展，并讨论其当前和将来的应用。虽然蛋白质组学通常集中于测量相对蛋白质浓度，但TPP提供了一种新颖的方法来收集有关表达数据集中不存在的蛋白质稳定性的补充信息。因此，该策略不仅在药物发现方面具有巨大潜力，而且在回答基本生物学问题方面也具有巨大潜力。