Experimental validation of free-energy-landscape reconstruction from non-equilibrium single-molecule force spectroscopy measurements (original) (raw)

Free-energy-landscape formalisms provide the fundamental conceptual framework for physical descriptions of how proteins and nucleic acids fold into specific three-dimensional structures. Although folding landscapes are difficult to measure experimentally, recent theoretical work by Hummer and Szabo has shown that landscape profiles can be reconstructed from non-equilibrium single-molecule force spectroscopy measurements using an extension of the Jarzynski equality. This method has been applied to simulations and experiments but never validated experimentally. We tested it using force-extension measurements on DNA hairpins with distinct, sequence-dependent folding landscapes. Quantitative agreement was found between the landscape profiles obtained from the non-equilibrium reconstruction and those from equilibrium probability distributions. We also tested the method on a riboswitch aptamer with three partially folded intermediate states, successfully reconstructing the landscape but f...