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Verification of the Crooks fluctuation theorem and recovery of RNA folding free energies

Verification of the Crooks fluctuation theorem and recovery of RNA folding free energies Atomic force microscopes and optical tweezers are widely used to probe the mechanical properties of individual molecules and molecular interactions, by exerting mechanical forces that induce transitions such as unfolding or dissociation. These transitions often occur under nonequilibrium conditions and are associated with hysteresis effects—features usually taken to preclude the extraction of equilibrium information from the experimental data. But fluctuation theorems 1,2,3,4,5 allow us to relate the work along nonequilibrium trajectories to thermodynamic free-energy differences. They have been shown to be applicable to single-molecule force measurements 6 and have already provided information on the folding free energy of a RNA hairpin 7,8 . Here we show that the Crooks fluctuation theorem 9 can be used to determine folding free energies for folding and unfolding processes occurring in weak as well as strong nonequilibrium regimes, thereby providing a test of its validity under such conditions. We use optical tweezers 10 to measure repeatedly the mechanical work associated with the unfolding and refolding of a small RNA hairpin 11 and an RNA three-helix junction 12 . The resultant work distributions are then analysed according to the theorem and allow us to determine the difference in folding free energy between an RNA molecule and a mutant differing only by one base pair, and the thermodynamic stabilizing effect of magnesium ions on the RNA structure. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Springer Journals

Verification of the Crooks fluctuation theorem and recovery of RNA folding free energies

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References (42)

Publisher
Springer Journals
Copyright
Copyright © 2005 by Nature Publishing Group
Subject
Science, Humanities and Social Sciences, multidisciplinary; Science, Humanities and Social Sciences, multidisciplinary; Science, multidisciplinary
ISSN
0028-0836
eISSN
1476-4687
DOI
10.1038/nature04061
Publisher site
See Article on Publisher Site

Abstract

Atomic force microscopes and optical tweezers are widely used to probe the mechanical properties of individual molecules and molecular interactions, by exerting mechanical forces that induce transitions such as unfolding or dissociation. These transitions often occur under nonequilibrium conditions and are associated with hysteresis effects—features usually taken to preclude the extraction of equilibrium information from the experimental data. But fluctuation theorems 1,2,3,4,5 allow us to relate the work along nonequilibrium trajectories to thermodynamic free-energy differences. They have been shown to be applicable to single-molecule force measurements 6 and have already provided information on the folding free energy of a RNA hairpin 7,8 . Here we show that the Crooks fluctuation theorem 9 can be used to determine folding free energies for folding and unfolding processes occurring in weak as well as strong nonequilibrium regimes, thereby providing a test of its validity under such conditions. We use optical tweezers 10 to measure repeatedly the mechanical work associated with the unfolding and refolding of a small RNA hairpin 11 and an RNA three-helix junction 12 . The resultant work distributions are then analysed according to the theorem and allow us to determine the difference in folding free energy between an RNA molecule and a mutant differing only by one base pair, and the thermodynamic stabilizing effect of magnesium ions on the RNA structure.

Journal

NatureSpringer Journals

Published: Sep 8, 2005

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