A comparative study of bioorthogonal reactions with azides
A comparative study of bioorthogonal reactions with azides
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DOI：
10.1021/cb6003228
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发表时间：
2006-01-01
影响因子：
4
通讯作者：
Bertozzi, Carolyn R.
中科院分区：
文献类型：
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作者：
Agard, Nicholas J.;Baskin, Jeremy M.;Bertozzi, Carolyn R.
代谢物和翻译后修饰物的检测。可以使用叠氮化物作为生物正交化学报告物来获得阳离子。一旦引入目标生物分子，无论是代谢或通过化学修饰。阳离子，叠氮化物可以使用三种高选择性反应之一用探针标记：Staudinger连接，Cu（I）催化的叠氮化物-炔环加成或应变促进的[3 + 2]环加成。在这里，我们在各种生物应用的背景下比较了这些化学，包括在复杂的裂解物和活细胞表面上标记生物分子。发现Cu（I）催化的反应对于检测蛋白质样品中的叠氮化物是最有效的，但由于试剂的毒性而与活细胞不相容。施陶丁格连接和使用优化的环辛炔的应变促进的[3 + 2]环加成对于在活细胞上标记叠氮化物是有效的。用于该应用的最佳试剂取决于叠氮化物的具体结构。这些结果为生物学家选择合适的连接化学提供了指导。
Detection of metabolites and post-translational modi. cations can be achieved using the azide as a bioorthogonal chemical reporter. Once introduced into target biomolecules, either metabolically or through chemical modi. cation, the azide can be tagged with probes using one of three highly selective reactions: the Staudinger ligation, the Cu(I)-catalyzed azide-alkyne cycloaddition, or the strain-promoted [3 + 2] cycloaddition. Here, we compared these chemistries in the context of various biological applications, including labeling of biomolecules in complex lysates and on live cell surfaces. The Cu(l)-catalyzed reaction was found to be most efficient for detecting azides in protein samples but was not compatible with live cells due to the toxicity of the reagents. Both the Staudinger ligation and the strain-promoted [3 + 2] cycloaddition using optimized cyclooctynes were effective for tagging azides on live cells. The best reagent for this application was dependent upon the specific structure of the azide. These results provide a guide for biologists in choosing a suitable ligation chemistry.