Regensburg 2016 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 44: Biotechnology & Bioengineering
BP 44.4: Vortrag
Mittwoch, 9. März 2016, 16:00–16:15, H45
Study of Reaction Networks with High-Throughput Nanoliter Thermophoresis — •Ferdinand Greiss1, Franziska Kriegel2, and Dieter Braun1 — 1Systems Biophysics, Quantitative Biosciences Munich (QBM), LMU, Munich, Germany — 2Molecular Biophysics, LMU, Munich, Germany
Quantifying the cooperative effect of protein binding is important and a well-established field in biochemistry. For instance, the binding of oxygen to hemoglobin is a widely known and thoroughly studied case of a positive homotropic binding reaction. This means that oxygen together with other oxygen molecules is binding in a positive cooperative manner. Many biological relevant reaction networks, e.g. transcription factors binding to DNA, include heterotropic binding interactions, both being negative or positive as well as with weak or strong cooperative effects.
We use synthetic DNA constructs as a simplified testbed to study the cooperative effects in heterotropic reaction networks with micro-scale thermophoresis (MST). All three DNA species have two different binding sites that can only access one partner. The advantage of the assay is that only one species needs to be labeled. The binding reaction network can be studied by independent single-point mutations, both in experiment and by theory.
With a newly designed high-throughput micro-scale thermophoresis setup, we are able to sample the large concentration space in a rapid, robust and user-friendly way.