Gallastegui Nerea, Groll Michael
Department of Biochemistry, Technische Universität München, Garching, Germany.
Methods Mol Biol. 2012;832:373-90. doi: 10.1007/978-1-61779-474-2_26.
The combination of X-ray crystallography and kinetic studies of proteasome:ligand complexes has proven to be an important tool in inhibitor analysis of this crucial protein degradation machinery. Here, we describe in detail the purification protocols, proteolytic activity assays, crystallisation methods, and structure determination for the yeast 20S proteasome (CP) in complex with its inhibitors. The fusion of these advanced techniques offers the opportunity to further optimise drugs which are already tested in different clinical phase studies, as well as to design new promising proteasome lead structures which might be suitable for their application in medicine, plant protection, and antibiotics.
X射线晶体学与蛋白酶体：配体复合物的动力学研究相结合，已被证明是分析这一关键蛋白质降解机制抑制剂的重要工具。在此，我们详细描述了与抑制剂结合的酵母20S蛋白酶体（CP）的纯化方案、蛋白水解活性测定、结晶方法和结构测定。这些先进技术的融合为进一步优化已在不同临床阶段研究中进行测试的药物提供了机会，同时也为设计新的有前景的蛋白酶体先导结构提供了机会，这些结构可能适用于医学、植物保护和抗生素领域。
