热蛋白质组学分析：一种全面评估蛋白质状态的技术

Thermal proteome profiling: a technique for a comprehensive assessment of protein status

摘要:

热蛋白质组学分析（thermal proteome profiling，TPP）是细胞热漂移测定（cellular thermal shift assay，CETSA）与定量质谱（quantitative mass spectrometry，MS）的结合，所以也称为MS-CETSA。热蛋白质组学分析通过测量不同加热温度下细胞或细胞裂解物中可溶蛋白的含量来确定整个蛋白质组的稳定性。蛋白质可以在与药物或代谢物等小分子、核酸或其他蛋白质相互作用或在翻译后修饰时改变其热稳定性，而热蛋白质组学分析可以根据有无配体结合蛋白质的热稳定性差异来确定靶蛋白。目前热蛋白质组学分析已成功应用于识别药物的靶点和脱靶点，探究蛋白质-代谢物和蛋白质-蛋白质的相互作用。总体上，国内对这个技术的了解仍然欠缺，对此，文中对热蛋白质组学分析的原理、方法、应用以及优势与局限性进行了综述。

Abstract:

Thermal proteome profiling (TPP) is a combination of cellular thermal shift assay (CETSA) and quantitative mass spectrometry (MS), also termed as MS-CETSA. TPP determines the stability of the entire proteome by measuring the content of soluble proteins in cells or cell lysates at different heating temperatures. Proteins can change their thermostability when interacting with small molecules (e.g., drugs or metabolites), nucleic acids, or other proteins or posttranslational modification, while TPP can identify target proteins based on the difference in thermostability with or without ligand-binding. At present, TPP has been applied to identify the targets and off-targets of drugs and interrogate protein-metabolite and protein-protein interactions. Due to limited understanding of this technology, this review introduced the principles, methods, applications, advantages and limitations of TPP.