Syntheses and Analyses of Modified Zif268 Zinc Finger Domains

In the current post-genomic research it is of high interest to gain deeper knowledge of the course of events leading to genetic diseases. Possessing the ability to already interfere at an early stage in the regulation of cellular processes is of special interest. Since the gene regulation is mainly accomplished by the interaction of DNA and proteins exhibiting sequence specific DNA binding abilities, the research is heading towards the development of modified DNA binders.One of the most prominent DNA binding domains in the human organism is the zinc finger motif.The main focus of this work lies on introducing additional metal binding sites into the entire sequence of the Zif268 zinc finger domain. Hence, a composition of a sequence selective DNA binding polypeptide including a possible functionality for example DNA cleavage is facilitated.The expressed protein ligation (EPL) approach enables the synthesis of a large polypeptide while keeping the possibility of artificial modifications.The results presented in this work show the effectiveness of a semi-synthetic approach which facilitates for example the introduction of additional metal binding sites into a selective DNA binding domain without losing its overall function analysed by different spectroscopic methods.