title: DNA origami as biocompatible surface to match single-molecule and ensemble experiments.
abstract: Single-molecule experiments on immobilized molecules allow unique insights into the dynamics of molecular machines and enzymes as well as their interactions. The immobilization, however, can invoke perturbation to the activity of biomolecules causing incongruities between single molecule and ensemble measurements. Here we introduce the recently developed DNA origami as a platform to transfer ensemble assays to the immobilized single molecule level without changing the nano-environment of the biomolecules. The idea is a stepwise transfer of common functional assays first to the surface of a DNA origami, which can be checked at the ensemble level, and then to the microscope glass slide for single-molecule inquiry using the DNA origami as a transfer platform. We studied the structural flexibility of a DNA Holliday junction and the TATA-binding protein (TBP)-induced bending of DNA both on freely diffusing molecules and attached to the origami structure by fluorescence resonance energy transfer. This resulted in highly congruent data sets demonstrating that the DNA origami does not influence the functionality of the biomolecule. Single-molecule data collected from surface-immobilized biomolecule-loaded DNA origami are in very good agreement with data from solution measurements supporting the fact that the DNA origami can be used as biocompatible surface in many fluorescence-based measurements.
authors.affiliation: Physikalische und Theoretische Chemie - NanoBioSciences, Technische Universität Braunschweig, Hans-Sommer-Strasse 10, 38106 Braunschweig, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Gietl
authors.forename: Phil
authors.initials: P
authors.lastname: Holzmeister
authors.forename: Dina
authors.initials: D
authors.lastname: Grohmann
authors.forename: Philip
authors.initials: P
authors.lastname: Tinnefeld
mesh_terms: D004247:DNA / Q000737:chemistry*; D045566:DNA, Cruciform / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D049329:Nanostructures / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D016385:TATA Box; D035181:TATA-Box Binding Protein / Q000378:metabolism
chemical_list: D045566:DNA, Cruciform; D035181:TATA-Box Binding Protein; D004247:DNA