Development and application of site specific fluorescent labeling of proteins

我们开发了用于标记 His 标签 ((His)_6) 的“开启”荧光探针，但后来发现它存在一些问题。本研究的目的是澄清和解决这些问题，开发荧光标记蛋白质的有用方法。我们的探针的主要问题是与没有 N 末端伯氨基的 His 标签的结合较弱。我们确定问题主要是由于组氨酸标签识别位点（一种金属复合物）造成的。我们探索了更好的 His 标签识别位点，并鉴定了一种金属络合物，它可以在任何位置与 His 标签结合。

We have developed "turn-on" fluorescent probes for labeling of His-tag ((His)_6), which later turned out to have some problems. The purpose of this research is the clarification and solution of these problems, to develop useful methodology for fluorescently label proteins. The main problem of our probes is the weak binding to His-tag without N-terminal primary amino group. We determined the problem is mainly due to the His-tag recognition site, a metal complex. We explored the better His-tag recognition site and identified a metal-complex, which can bind to His-tag at any position.