Sinz Andrea
Biotechnological-Biomedical Center, Faculty of Chemistry and Mineralogy, University of Leipzig, D-04103 Leipzig, Germany.
Mass Spectrom Rev. 2006 Jul-Aug;25(4):663-82. doi: 10.1002/mas.20082.
Closely related to studying the function of a protein is the analysis of its three-dimensional structure and the identification of interaction sites with its binding partners. An alternative approach to the high-resolution methods for three-dimensional protein structure analysis, such as X-ray crystallography and NMR spectroscopy, consists of covalently connecting two functional groups of the protein(s) under investigation. The location of the created cross-links imposes a distance constraint on the location of the respective side chains and allows one to draw conclusions on the three-dimensional structure of the protein or a protein complex. Recently, chemical cross-linking of proteins has been combined with a mass spectrometric analysis of the created cross-linked products. This review article describes the most popular cross-linking reagents for protein structure analysis and gives an overview of the different available strategies that employ chemical cross-linking and different mass spectrometric techniques. The challenges for mass spectrometry caused by the enormous complexity of the cross-linking reaction mixtures are emphasized. The various approaches described in the literature to facilitate the mass spectrometric detection of cross-linked products as well as computer software for data analyses are reviewed.
与研究蛋白质功能密切相关的是对其三维结构的分析以及对其与结合伙伴相互作用位点的鉴定。用于三维蛋白质结构分析的高分辨率方法，如X射线晶体学和核磁共振光谱学，有一种替代方法，即共价连接被研究蛋白质的两个官能团。所形成交联的位置对相应侧链的位置施加了距离限制，从而使人们能够对蛋白质或蛋白质复合物的三维结构得出结论。最近，蛋白质的化学交联已与对所形成交联产物的质谱分析相结合。这篇综述文章描述了用于蛋白质结构分析的最常用交联试剂，并概述了采用化学交联和不同质谱技术的不同可用策略。强调了交联反应混合物的巨大复杂性给质谱分析带来的挑战。综述了文献中描述的促进交联产物质谱检测的各种方法以及用于数据分析的计算机软件。
