Uta Wienken (née Steinbach)

German

Surface receptors and their interactions are in focus of biomedical research as they play a fundamental role in signal transduction and are involved in cancer development and progression.
In my PhD thesis I apply the Molecular Force Assay (MFA) on living cells to investigate the binding properties of such receptors in their physiological environment. The method relies on the well-established principle of the molecular force assay (MFA) that was so far only applied on molecules immobilized on surfaces. It is an ensemble measurement that analyzes binding forces of receptor ligand interactions on a single molecule level by directly comparing the binding force of interest to a known reference interaction. Reference and probe are clamped in series between two surfaces. When separating the surfaces, the molecular complex with the weaker bond is more likely to rupture. A fluorescent label at the reference complex indicates the outcome of the experiment. This highly parallel method ensures to measure specific binding of a cell surface receptor to its ligand by exertion of a certain reference force that exceeds the strength of unspecific interactions. Thus, the introduced assay is a promising method that is not only capable of screening for various receptors on the membrane of a certain cell line in one experiment but also information about the mechanical properties of those interactions.

(under my maiden name Uta Steinbach:)
D. Ho, J. L. Zimmermann, F. A. Dehmelt, U. Steinbach, M. Erdmann, P. Severin, K. Falter, H. E. Gaub:
"Force-driven separation of short double stranded DNA"
Biophysical Journal. 97:3158-3167 (2009)

J. Morfill, J. Neumann, K. Blank, U. Steinbach, E. M. Puchner, K. E. Gottschalk, and H. E. Gaub.
"Force-based analysis of multidimensional energy landscapes: application of dynamic force spectroscopy and steered molecular dynamics simulations to an antibody fragment-peptide complex"
J Mol Biol 381(5):1253-1266 (2008)