Synthesis of polystyrene microspheres and functionalization with Pd0 nanoparticles to perform bioorthogonal organometallic chemistry in living cells
Synthesis of polystyrene microspheres and functionalization with Pd0 nanoparticles to perform bioorthogonal organometallic chemistry in living cells
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DOI：
10.1038/nprot.2012.052
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发表时间：
2012-06-01
期刊：
影响因子：
14.8
通讯作者：
Bradley, Mark
中科院分区：
文献类型：
--
作者：
Unciti-Broceta, Asier;Johansson, Emma M. V.;Bradley, Mark
我们开发了小型化异质 Pd-0 催化剂（Pd-0 微球），能够进入细胞、无害地停留在细胞质内并介导有效的生物正交有机金属化学反应（例如，烯丙基氨基甲酸酯裂解和铃木-宫浦交叉偶联）。这种方法是对可用于在细胞内进行化学反应的工具箱的主要补充。在这里，我们描述了从现成的起始材料合成 Pd-0-微球的完整方案（通过合成尺寸控制的氨基功能化聚苯乙烯微球），以及它们的表征（电子显微镜和钯定量）和功能验证（“溶液中”和“细胞质中”转换）。催化装置从开始合成到功能评价需要5天的工作时间。
We have developed miniaturized heterogeneous Pd-0-catalysts (Pd-0-microspheres) with the ability to enter cells, stay harmlessly within the cytosol and mediate efficient bioorthogonal organometallic chemistries (e.g., allylcarbamate cleavage and Suzuki-Miyaura cross-coupling). This approach is a major addition to the toolbox available for performing chemical reactions within cells. Here we describe a full protocol for the synthesis of the Pd-0-microspheres from readily available starting materials (by the synthesis of size-controlled amino-functionalized polystyrene microspheres), as well as for their characterization (electron microscopy and palladium quantitation) and functional validation ('in solution' and 'in cytoplasm' conversions). From the beginning of the synthesis to functional evaluation of the catalytic device requires 5 d of work.