The aim of these studies is the quantitative investigation of protein-protein interactions in the AP1 system in vivo. First results of FCS measurements show an exchange in the nucleus of the proteins Fos-CFP and Jun-YFP in the stably mono-transfected HeLa-Cells. This is also shown by fitting the bleaching curves measured in the nucleus with an appropriate model. We obtained dissociation times between 10 and 20 seconds in the nucleus. In the autocorrelation function a free and an obstructed component of diffusion are shown. For further studies doubly transfected cells with both proteins, Fos-CFP and Jun-YFP, were prepared. These cells will now be characterized with FCCS to investigate the protein-protein interactions. In order to obtain the dissociation rates of the complex in the cell nucleus bleaching curves will be recorded on these cell lines. We also overexpressed and purified Jun-YFP and Fos-CFP for in vitro studies.

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hdl.handle.net/1765/77487
5th Graduate Students Meeting of the German Cancer Research Center (DKFZ)
Biophysical Genomics, Department Cell Biology & Genetics

Baudendistel, N., Knoch, T., Müller, G., Wachsmuth, M., Weidemann, T., Waldeck, W., & Langowski, J. (2002). In vivo characterization of protein-protein interactions in the AP1 system with fluorescence correlation spectroscopy (FCS).. Presented at the 5th Graduate Students Meeting of the German Cancer Research Center (DKFZ). Retrieved from http://hdl.handle.net/1765/77487