使用溶液 NMR 光谱法鉴定蛋白质中的药物结合位点的方案。
Protocol to identify drug-binding sites in proteins using solution NMR spectroscopy.
机构信息
Department of Biochemistry, Case Western Reserve University, Cleveland, OH 44106, USA.
Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO 80045, USA.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101842. doi: 10.1016/j.xpro.2022.101842. Epub 2022 Nov 11.
Dusquetide is a next-generation IDR (innate defense regulator) targeting the major autophagy receptor protein SQSTM1/p62 and modulating the innate immune response. Here, we describe a protocol for determining dusquetide-binding sites of p62 by solution NMR spectroscopy. Step-by-step technique details were provided, including sample preparation, NMR experiment setup, data processing, and binding site analysis. This protocol could be applied to characterize other small molecules targeting the ZZ domain of p62 (9 kDa) or other proteins containing ZZ domains. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).
杜斯奎肽是一种针对主要自噬受体蛋白 SQSTM1/p62 的下一代 IDR（固有防御调节剂），可调节固有免疫反应。在这里，我们描述了一种通过溶液 NMR 光谱法确定 p62 与杜斯奎肽结合位点的方案。提供了详细的分步技术细节，包括样品制备、NMR 实验设置、数据处理和结合位点分析。该方案可用于表征其他针对 p62（9 kDa）的 ZZ 结构域的小分子或其他含有 ZZ 结构域的蛋白质。有关此方案使用和执行的完整详细信息，请参阅 Zhang 等人。（2022 年）。
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