Copper-free click chemistry for dynamic in vivo imaging
Copper-free click chemistry for dynamic in vivo imaging
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DOI：
10.1073/pnas.0707090104
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发表时间：
2007-10-23
影响因子：
11.1
通讯作者：
Bertozzi, Carolyn R.
中科院分区：
文献类型：
--
作者：
Baskin, Jeremy M.;Prescher, Jennifer A.;Bertozzi, Carolyn R.
活细胞中蛋白质的动态成像通常是通过使用遗传编码的报告进行的，这种方法不能扩展到其他类别的生物分子，如多糖和脂类。在这里，我们报告了一种无铜点击化学的变体，它可以在生命系统中快速和选择性地标记这些生物分子，克服了典型的铜催化反应的内在毒性。关键试剂是取代的环辛炔，它具有环张力和吸电子的氟取代基，它们共同促进[3+2]偶极环加成反应，叠氮化合物以代谢方式安装到生物分子中。这种无铜的点击反应具有与铜催化的反应类似的动力学，在活细胞上几分钟内进行，没有明显的毒性。利用这项技术，我们研究了糖链转运的动力学，并确定了一组唾液酸糖共轭化合物具有出人意料的快速内化动力学。
Dynamic imaging of proteins in live cells is routinely performed by using genetically encoded reporters, an approach that cannot be extended to other classes of biomolecules such as glycans and lipids. Here, we report a Cu-free variant of click chemistry that can label these biomolecules rapidly and selectively in living systems, overcoming the intrinsic toxicity of the canonical Cu-catalyzed reaction. The critical reagent, a substituted cyclooctyne, possesses ring strain and electron-withdrawing fluorine substituents that together promote the [3 + 2] dipolar cycloaddition with azides installed metabolically into biornolecules. This Cu-free click reaction possesses comparable kinetics to the Cu-catalyzed reaction and proceeds within minutes on live cells with no apparent toxicity. With this technique, we studied the dynamics of glycan trafficking and identified a population of sialoglycoconjugates with unexpectedly rapid internalization kinetics.