Development and application of site specific fluorescent labeling of proteins

我们已经开发了用于标记His标签（（His）_6）的"开启"荧光探针，后来发现存在一些问题。本研究的目的是澄清和解决这些问题，发展有用的荧光标记蛋白质的方法。我们的探针的主要问题是与没有N-末端伯氨基的His-标签的弱结合。我们确定这个问题主要是由于His标签识别位点，一种金属复合物。我们探索了更好的His标签识别位点，并鉴定了一种金属复合物，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.