Berlin 2012 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 7: Posters: Proteins
BP 7.23: Poster
Montag, 26. März 2012, 17:30–19:30, Poster A
Investigation of the PhoB-Interaction with the DNA - minor groove by Single Molecule Force Spectroscopy — •Adeline Bieker1, Volker Walhorn1, Gesa Niemann2, Markus Ritzefeld2, Norbert Sewald2, and Dario Anselmetti1 — 1Experimental Biophysics and Applied Nanoscience, University of Bielefeld, Germany — 2Organic and Bioorganic Chemistry, University of Bielefeld, Germany
Interactions between proteins and DNA are essential for the regulation of cellular processes in all living organisms. In this context, it is of special interest to investigate and quantify the sequence-specific molecular recognition between transcription factors and their cognate DNA sequences [1].
We investigated point mutated proteins originating from the DNA-binding domain (DBD) of the Escherichia coli transcription factor PhoB. By means of AFM-based Single Molecule Force Spectroscopy (SMFS) we acquired binding forces and molecular elasticities to elucidate the complex stability. Based on the Bell-Evans-Modell [2] we estimated the thermal dissociation rate constants (koff) and the molecular interaction length (xβ), that allowed a structure related interpretation of the physical binding mechanisms involved.
[1] Small. 2009 Apr;5(4):484-495.
[2] Biophys. J. 1997;72:1541-1555.