Comparative energy measurements in single molecule interactions.

Single molecule experiments have opened promising new avenues of investigations in biology, but the quantitative interpretation of results remains challenging. In particular, there is a need for a comparison of such experiments with theoretical methods. We experimentally determine the activation free energy for single molecule interactions between two synaptic proteins syntaxin 1A and synaptobrevin 2, using an atomic force microscope and the Jarzynski equality of nonequilibrium thermodynamics. The value obtained is shown to be reasonably consistent with that from single molecule reaction rate theory. The temperature dependence of the spontaneous dissociation lifetime along with different pulling speeds is used to confirm the approach to the adiabatic limit. This comparison of the Jarzynski equality for intermolecular interactions extends the procedure for calculation of activation energies in nonequilibrium processes.

[1]  R. Scheller,et al.  Mechanisms of synaptic vesicle exocytosis. , 2000, Annual review of cell and developmental biology.

[2]  R. Merkel Force spectroscopy on single passive biomolecules and single biomolecular bonds , 2001 .

[3]  T. Südhof,et al.  Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly. , 1994, The EMBO journal.

[4]  R. Swendsen,et al.  Comparison of free energy methods for molecular systems. , 2006, The Journal of chemical physics.

[5]  U. Mohideen,et al.  Fiber optic interferometry for precision measurement of the voltage and frequency dependence of the displacement of piezoelectric tubes , 2001 .

[6]  Klaus Schulten,et al.  Mechanical unfolding intermediates in titin modules , 1999, Nature.

[7]  M. Hegner,et al.  Temperature dependence of unbinding forces between complementary DNA strands. , 2002, Biophysical journal.

[8]  H. Gaub,et al.  Adhesion forces between individual ligand-receptor pairs. , 1994, Science.

[9]  Reinhard Jahn,et al.  Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 Å resolution , 1998, Nature.

[10]  J. Liphardt,et al.  Reversible Unfolding of Single RNA Molecules by Mechanical Force , 2001, Science.

[11]  P K Hansma,et al.  Stepwise unfolding of titin under force-clamp atomic force microscopy. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[12]  G. Hummer,et al.  Free energy reconstruction from nonequilibrium single-molecule pulling experiments , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Thomas B Woolf,et al.  Theory of a systematic computational error in free energy differences. , 2002, Physical review letters.

[14]  E. Evans Probing the relation between force--lifetime--and chemistry in single molecular bonds. , 2001, Annual review of biophysics and biomolecular structure.

[15]  K. Schulten,et al.  Molecular dynamics study of unbinding of the avidin-biotin complex. , 1997, Biophysical journal.

[16]  G. I. Bell Models for the specific adhesion of cells to cells. , 1978, Science.

[17]  P. Steiner,et al.  Interactions between synaptic vesicle fusion proteins explored by atomic force microscopy , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Michael R. Shirts,et al.  Equilibrium free energies from nonequilibrium measurements using maximum-likelihood methods. , 2003, Physical review letters.

[19]  Gerhard Hummer,et al.  Intrinsic rates and activation free energies from single-molecule pulling experiments. , 2006, Physical review letters.

[20]  C. Kiang,et al.  Experimental free energy surface reconstruction from single-molecule force spectroscopy using Jarzynski's equality. , 2007, Physical review letters.

[21]  J. Klafter,et al.  Beyond the conventional description of dynamic force spectroscopy of adhesion bonds , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  G. Crooks Path-ensemble averages in systems driven far from equilibrium , 1999, cond-mat/9908420.

[23]  C. Bustamante,et al.  Overstretching B-DNA: The Elastic Response of Individual Double-Stranded and Single-Stranded DNA Molecules , 1996, Science.

[24]  C. Jarzynski Nonequilibrium Equality for Free Energy Differences , 1996, cond-mat/9610209.

[25]  C. Jarzynski Rare events and the convergence of exponentially averaged work values. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[26]  G. Crooks Entropy production fluctuation theorem and the nonequilibrium work relation for free energy differences. , 1999, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[27]  C. Jarzynski,et al.  A “fast growth” method of computing free energy differences , 2001 .

[28]  V. Parpura,et al.  Single molecule mechanical probing of the SNARE protein interactions. , 2006, Biophysical journal.

[29]  A. Brunger,et al.  Single molecule observation of liposome-bilayer fusion thermally induced by soluble N-ethyl maleimide sensitive-factor attachment protein receptors (SNAREs). , 2004, Biophysical journal.

[30]  I. Tinoco,et al.  Equilibrium Information from Nonequilibrium Measurements in an Experimental Test of Jarzynski's Equality , 2002, Science.

[31]  J. Bechhoefer,et al.  Calibration of atomic‐force microscope tips , 1993 .

[32]  H. Gaub,et al.  Unfolding pathways of individual bacteriorhodopsins. , 2000, Science.

[33]  Axel T. Brunger,et al.  Single-molecule studies of SNARE complex assembly reveal parallel and antiparallel configurations , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Nonlinear elasticity of an alpha -helical polypeptide. , 2004, Physical review. E, Statistical, nonlinear, and soft matter physics.

[35]  Precision measurement of the Casimir force using gold surfaces , 2000, quant-ph/0005088.

[36]  C. Jarzynski,et al.  Verification of the Crooks fluctuation theorem and recovery of RNA folding free energies , 2005, Nature.