Department of Biochemistry, 5323 Harry Hines Boulevard, University of Texas Southwestern Medical Center, Dallas, TX 75390-9038, USA.
Curr Opin Chem Biol. 2013 Feb;17(1):90-101. doi: 10.1016/j.cbpa.2012.10.034. Epub 2012 Nov 10.
Photocrosslinking approaches can be used to map interactome networks within the context of living cells. Photocrosslinking methods rely on use of metabolic engineering or genetic code expansion to incorporate photocrosslinking analogs of amino acids or sugars into cellular biomolecules. Immunological and mass spectrometry techniques are used to analyze crosslinked complexes, thereby defining specific interactomes. Because photocrosslinking can be conducted in native, cellular settings, it can be used to define context-dependent interactions. Photocrosslinking methods are also ideally suited for determining interactome dynamics, mapping interaction interfaces, and identifying transient interactions in which intrinsically disordered proteins and glycoproteins engage. Here we discuss the application of cell-based photocrosslinking to the study of specific problems in immune cell signaling, transcription, membrane protein dynamics, nucleocytoplasmic transport, and chaperone-assisted protein folding.
光交联方法可用于在活细胞的背景下绘制相互作用组网络。光交联方法依赖于代谢工程或遗传密码扩展，将氨基酸或糖的光交联类似物掺入细胞生物分子中。免疫和质谱技术用于分析交联复合物，从而定义特定的相互作用组。由于光交联可以在天然的细胞环境中进行，因此它可用于定义依赖于上下文的相互作用。光交联方法也非常适合于确定相互作用组的动态、映射相互作用界面以及识别固有无序蛋白质和糖蛋白参与的瞬时相互作用。在这里，我们讨论了基于细胞的光交联在免疫细胞信号转导、转录、膜蛋白动力学、核质转运和伴侣辅助蛋白折叠等特定问题研究中的应用。
