Center for Integrated Protein Science Munich (CIPS(M)), Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany.
ChemMedChem. 2012 Jul;7(7):1146-59. doi: 10.1002/cmdc.201200057. Epub 2012 Mar 19.
Proteases are important targets for the treatment of human disease. Several protease inhibitors have failed in clinical trials due to a lack of in vivo specificity, indicating the need for studies of protease function and inhibition in complex, disease-related models. The tight post-translational regulation of protease activity complicates protease analysis by traditional proteomics methods. Activity-based protein profiling is a powerful technique that can resolve this issue. It uses small-molecule tools-activity-based probes-to label and analyze active enzymes in lysates, cells, and whole animals. Over the last twelve years, a wide variety of protease activity-based probes have been developed. These synthetic efforts have enabled techniques ranging from real-time in vivo imaging of protease activity to high-throughput screening of uncharacterized proteases. This Review introduces the general principles of activity-based protein profiling and describes the recent advancements in probe design and analysis techniques, which have increased the knowledge of protease biology and will aid future protease drug discovery.
蛋白酶是治疗人类疾病的重要靶点。由于缺乏体内特异性，几种蛋白酶抑制剂在临床试验中失败，这表明需要在复杂的、与疾病相关的模型中研究蛋白酶的功能和抑制作用。蛋白酶活性的严格翻译后调控使得传统蛋白质组学方法对蛋白酶的分析变得复杂。基于活性的蛋白质谱分析是一种强大的技术，可以解决这个问题。它使用小分子工具——活性探针，来标记和分析裂解物、细胞和整个动物中的活性酶。在过去的十二年中，已经开发出了各种各样的蛋白酶活性探针。这些合成工作使从实时体内蛋白酶活性成像到未表征蛋白酶的高通量筛选等技术成为可能。本文综述了基于活性的蛋白质谱分析的一般原理，并描述了探针设计和分析技术的最新进展，这些进展增加了对蛋白酶生物学的认识，并将有助于未来的蛋白酶药物发现。
