Tri-Institutional PhD Program in Chemical Biology, New York, NY, USA.
Chemical Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
Methods Mol Biol. 2020;2133:201-219. doi: 10.1007/978-1-0716-0434-2_10.
The development of expressed protein ligation (EPL) widened the scope of questions that could be addressed by mechanistic biochemistry. Protein trans-splicing (PTS) relies on the same basic chemical principles, but utilizes split inteins to tracelessly ligate distinct peptide or polypeptide fragments together with native peptide bonds. Here we present a method to adapt PTS methodologies for their use in live cells, in order to deliver synthetic or native histone modifications. As an example, we provide a protocol to incorporate a small molecule fluorophore into chromatinized histones. The protocol should be easily adaptable to incorporate other modifications to chromatin in vivo.
表达蛋白连接（EPL）的发展拓宽了可以通过机制生物化学解决的问题范围。蛋白质转剪接（PTS）依赖于相同的基本化学原理，但利用分裂的内含肽无痕地将不同的肽或多肽片段与天然肽键连接在一起。在这里，我们提出了一种将 PTS 方法应用于活细胞的方法，以便递送合成或天然组蛋白修饰。作为一个例子，我们提供了一个将小分子荧光团掺入组蛋白的方案。该方案应该很容易适应于在体内将其他修饰物掺入到染色质中。
