Topics of research:
I am investigating the interaction between small molecule ligands and the cyclic nucleotide binding domain (CNBD) of human hyperpolarization and cyclic nucleotide gated ion channels (HCN). The aim of my research project is the development of a ligand binding assay based on the surface expression of the C-Linker-CNBD- domain of the HCN4 channel as a fusion protein. Therefore, I use the Autodisplay technique, which enables the surface display of recombinant proteins on the outer membrane surface of E. coli cells.
The fluorescence intensity of 8-Fluo-cAMP bound to the displayed protein is measured by flow cytometry. A competitive binding assay should be the basis for a quick and easy screen for potential ligands. The procedure will be transferred to other HCN-C-Linker-CNBD subtypes. In this way, a specific testing of ligands can be carried out.