Chemistry for the analysis of protein-protein interactions: Rapid and efficient cross-linking triggered by long wavelength light
Chemistry for the analysis of protein-protein interactions: Rapid and efficient cross-linking triggered by long wavelength light
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DOI：
10.1073/pnas.96.11.6020
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发表时间：
1999-05-25
影响因子：
11.1
通讯作者：
Kodadek, T
中科院分区：
文献类型：
--
作者：
Fancy, DA;Kodadek, T
化学交联是一种潜在的有用的技术，用于探测多蛋白质复合物的结构。然而，使用典型的双官能交联剂的分析通常遭受低产率，并且亲核侧链的大规模修饰可导致归因于结构不稳定的人为结果。我们在这里报告从头设计和开发的一种类型的蛋白质交联反应，使用光生氧化剂介导的相关蛋白质的快速和有效的交联。该过程涉及在电子受体过硫酸铵和感兴趣的蛋白质的存在下，用可见光对三-联吡啶钌（II）二价阳离子进行简短的光解。交联产物的产率非常高，
Chemical cross-linking is a potentially useful technique for probing the architecture of multiprotein complexes. However, analyses using typical bifunctional cross-linkers often suffer from poor yields, and large-scale modification of nucleophilic side chains can result in artifactual results attributable to structural destabilization. We report here the de novo design and development of a type of protein cross-linking reaction that uses a photogenerated oxidant to mediate rapid and efficient cross-linking of associated proteins. The process involves brief photolysis of tris-bipyridylruthenium(II) dication with visible light in the presence of the electron acceptor ammonium persulfate and the proteins of interest. Very high yields of cross-linked products can be obtained with irradiation times of