Electron-tunneling data suggest that a noncovalently-bonded complex of three molecules, two recognition molecules that present hydrogen-bond donor and acceptor sites via a carboxamide group, and a DNA base, remains bound for seconds. This is surprising, given that imino-proton exchange rates show that basepairs in a DNA double helix open on millisecond timescales. The long lifetime of the three...

Exerting manual forces is critical during occupational performance. Therefore, being able to estimate maximum force capacity is particularly useful for determining how these manual exertion demands relate to available capacity. To facilitate this type of prediction requires a complete understanding of how maximum force capacity is governed biomechanically. This research focused on identifying h...

Zyxin is a focal adhesion protein that has been implicated in the modulation of cell adhesion and motility, and is hypothesized to be a mechano-sensor in integrin-mediated responses to mechanical force. To test the functional role of zyxin in the mechanotransduction of microvascular smooth muscle cells (VSMC), we utilized atomic force microscopy (AFM) to apply localized pulling forces to VSMC t...

We studied the velocity dependence of mechanical unfolding of single protein molecules with the atomic force microscope. We showed that with enough realizations, the free energy surfaces reconstructed from Jarzynski's equality converge with respect to pulling velocity, in good agreement with theory. Using the I27 domain of titin as an example, we estimated the required number of realizations fo...

Single-molecule manipulation techniques reveal that the mechanical resistance of a protein depends on the direction of the applied force. Using a lattice model of polymers, we show that changing the pulling direction leads to different phase diagrams. The simple model proposed here indicates that in one case the system undergoes a transition akin to the unzipping of a beta sheet, while in the o...

We study a class of micromanipulation experiments, exemplified by the pulling apart of the two strands of double-stranded DNA. When the pulling force is increased to a critical value, an "unzipping" transition occurs. For random DNA sequences with short-ranged correlations, we obtain exact results for the number of monomers liberated and the specific heat, including the critical behavior at the...

The active site of the Haloalkane Dehydrogenase (HaloTag) enzyme can be covalently attached to a chloroalkane ligand providing a mechanically strong tether, resistant to large pulling forces. Here we demonstrate the covalent tethering of protein L and I27 polyproteins between an atomic force microscopy (AFM) cantilever and a glass surface using HaloTag anchoring at one end and thiol chemistry a...

*Correspondence: [email protected] 1Laboratoire de Physique, Université de Lyon, Ecole Normale Supérieure de Lyon, CNRS, 46 Allée d’Italie, 69007 Lyon, France Abstract We report on an experimental device that makes it possible to assess the frictional properties of the contact between a slider and a horizontal surface, and to study the resulting trajectories of the slider whe...

Dynamic force spectroscopy has become indispensable for the exploration of the mechanical properties of proteins. In force-ramp experiments, performed by utilizing a time-dependent pulling force, the peak forces for unfolding transitions in a multimeric protein (D)(N) are used to map the free energy landscape for unfolding for a protein domain D. We show that theoretical modeling of unfolding t...

This article describes the development and the examination of surface coatings that suppress the adhesion between glass surfaces and polymer microspheres. Superparamagnetic doping allowed for exerting magnetic forces on the microbeads. The carboxyl functionalization of the polymer provided the means for coating the beads with polyethylene glycol (PEG) with different molecular weight. Under grav...