Dresden 2014 – scientific programme

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BP: Fachverband Biologische Physik

BP 2: DNA/RNA and related enzymes

BP 2.8: Talk

Monday, March 31, 2014, 11:45–12:00, ZEU 250

Hybrid Single-Molecule-FRET-Magnetic Tweezers Probe Force-Dependent Conformational Space — •Marko Swoboda, Maj Svea Grieb, Stephan Friebe, Steffen Hahn, and Michael Schlierf — B CUBE, TU Dresden, Arnoldstraße 18, 01307 Dresden, Germany

Single-molecule methods are often separated in force-based and fluorescence-based techniques. Single-molecule Förster resonance energy transfer (smFRET) has proven to be a powerful tool to probe distance changes on the nanometer scale. The technique therefore perfectly matches biomolecular dimensions but lacks the ability to manipulate molecules. Magnetic tweezers are a prime tool to study the mechanics of DNA and its interacting enzymes but their spatial resolution is limited to the direction of force and torque application.

Here, we demonstrate a hybrid instrument, combining smFRET and magnetic tweezers. This allows us to simultaneously probe conformational dynamics in a molecular system, using both force and distance information. We demonstrate the instrument’s capabilities by studying the behavior of a fluorescently labeled DNA undergoing rapid conformational fluctuations.