Gaczynska Maria, Osmulski Pawel A
Institute of Biotechnology, University of Texas Health Science Center at San Antonio, USA.
Methods Mol Biol. 2005;301:3-22. doi: 10.1385/1-59259-895-1:003.
The fast-track approval of a proteasome inhibitor, PS-341, to treat multiple myeloma spurred a wave of interest in both the proteasome itself and small-molecule compounds blocking its activities. Besides being candidates for drugs against cancer, autoimmune diseases, inflammation, or stroke, specific proteasome inhibitors are indispensable tools for biochemical and cell biology investigations of the proteasome and proteasome-ubiquitin system. Numerous synthetic peptide derivatives, such as boronates, epoxides, aldehydes, vinyl sulfones, cyclic peptides, and lactones, block the N-terminal threonine-type active centers of the enzyme, halting the cleavage of proteasomal protein substrates both in vitro and in vivo. Because some of the proteasomal inhibitors exhibit a high specificity toward only one particular type of an active center of the proteasome, they constitute valuable probes for testing the mechanism of proteolysis catalyzed by the enzyme. In this chapter we discuss the most common applications of available proteasome inhibitors. In addition to the best-known competitive inhibitors, we also describe the benefits from the use of allosteric inhibitors, which induce distinct but less understood in vitro and in vivo effects on the proteasomal machinery. Finally, we present the application of the basic biochemical procedures to decipher the mechanism of interactions of a novel compound with the proteasome.
蛋白酶体抑制剂PS - 341用于治疗多发性骨髓瘤的快速批准引发了人们对蛋白酶体本身以及阻断其活性的小分子化合物的广泛关注。除了作为抗癌、自身免疫性疾病、炎症或中风药物的候选物外，特定的蛋白酶体抑制剂还是蛋白酶体和蛋白酶体 - 泛素系统生化及细胞生物学研究中不可或缺的工具。众多合成肽衍生物，如硼酸盐、环氧化物、醛、乙烯基砜、环肽和内酯，可阻断该酶的N端苏氨酸型活性中心，在体外和体内均能阻止蛋白酶体蛋白底物的裂解。由于一些蛋白酶体抑制剂仅对蛋白酶体的一种特定活性中心表现出高特异性，它们构成了测试该酶催化蛋白水解机制的有价值探针。在本章中，我们将讨论现有蛋白酶体抑制剂的最常见应用。除了最知名的竞争性抑制剂外，我们还将描述使用变构抑制剂的益处，这些抑制剂对蛋白酶体机制在体外和体内会产生独特但尚不太清楚的影响。最后，我们将介绍基本生化程序在解读新型化合物与蛋白酶体相互作用机制方面的应用。
