Wu Ling, Xu Bin
Department of Biochemistry and Center for Drug Discovery, Virginia Polytechnic Institute and State University, Blacksburg, Virginia.
Curr Protoc Protein Sci. 2015 Feb 2;79:19.26.1-19.26.14. doi: 10.1002/0471140864.ps1926s79.
Photoaffinity cross-linking is a rapidly developing technology for studying biomolecular interactions, including protein ligand-receptor binding. This technology provides detailed binding information including receptor contact sites, active conformation of receptor-ligand complexes, global binding surfaces, and binding modes. Advancements in genetic technology have enabled non-natural photoactive amino acid derivatives to be incorporated into designer or target proteins, providing a host of new opportunities for manufacturing protein photo-probes while bypassing the traditional peptide or small protein limits of classical chemical synthesis. This unit provides several protocols for performing basic photoaffinity cross-linking and related analyses for applications in ligand-receptor binding and protein-protein interactions.
光亲和交联是一种快速发展的用于研究生物分子相互作用的技术,包括蛋白质配体-受体结合。该技术提供详细的结合信息,包括受体接触位点、受体-配体复合物的活性构象、整体结合表面和结合模式。基因技术的进步使得非天然光活性氨基酸衍生物能够被整合到设计蛋白或目标蛋白中,在绕过传统肽或经典化学合成的小蛋白限制的同时,为制造蛋白质光探针提供了大量新机会。本单元提供了几种用于进行基本光亲和交联及相关分析的方案,以应用于配体-受体结合和蛋白质-蛋白质相互作用研究。