A modular spring-loaded actuator for mechanical activation of membrane proteins.

How cells respond to mechanical forces by converting them into biological signals underlie crucial cellular processes. Our understanding of mechanotransduction has been hindered by technical barriers, including limitations in our ability to effectively apply low range piconewton forces to specific mechanoreceptors on cell membranes without laborious and repetitive trials. To overcome these challenges we introduce the Nano-winch, a robust, easily assembled, programmable DNA origami-based molecular actuator. The Nano-winch is designed to manipulate multiple mechanoreceptors in parallel by exerting fine-tuned, low- piconewton forces in autonomous and remotely activated modes via adjustable single- and double-stranded DNA linkages, respectively. Nano-winches in autonomous mode can land and operate on the cell surface. Targeting the device to integrin stimulated detectable downstream phosphorylation of focal adhesion kinase, an indication that Nano-winches can be applied to study cellular mechanical processes. Remote activation mode allowed finer extension control and greater force exertion. We united remotely activated Nano-winches with single-channel bilayer experiments to directly observe the opening of a channel by mechanical force in the force responsive gated channel protein, BtuB. This customizable origami provides an instrument-free approach that can be applied to control and explore a diversity of mechanotransduction circuits on living cells.

Mechanisms and Dynamics of the Bacterial Flagellar Motor.

Many bacteria are able to actively propel themselves through their complex environment, in search of resources and suitable niches. The source of this propulsion is the Bacterial Flagellar Motor (BFM), a molecular complex embedded in the bacterial membrane which rotates a flagellum. In this chapter we review the known physical mechanisms at work in the motor. The BFM shows a highly dynamic behavior in its power output, its structure, and in the stoichiometry of its components. Changes in speed, rotation direction, constituent protein conformations, and the number of constituent subunits are dynamically controlled in accordance to external chemical and mechanical cues. The mechano-sensitivity of the motor is likely related to the surface-sensing ability of bacteria, relevant in the initial stage of biofilm formation.

Correction: Kinetic analysis methods applied to single motor protein trajectories.

Correction for 'Kinetic analysis methods applied to single motor protein trajectories' by A. L. Nord et al., Phys. Chem. Chem. Phys., 2018, 20, 18775-18781.

Kinetic analysis methods applied to single motor protein trajectories.

Molecular motors convert chemical or electrical energy into mechanical displacement, either linear or rotary. Under ideal circumstances, single-molecule measurements can spatially and temporally resolve individual steps of the motor, revealing important properties of the underlying mechanochemical process. Unfortunately, steps are often hard to resolve, as they are masked by thermal noise. In such cases, details of the mechanochemistry can nonetheless be recovered by analyzing the fluctuations in the recorded traces. Here, we expand upon existing statistical analysis methods, providing two new avenues to extract the motor step size, the effective number of rate-limiting chemical states per translocation step, and the compliance of the link between the motor and the probe particle. We first demonstrate the power and limitations of these methods using simulated molecular motor trajectories, and we then apply these methods to experimental data of kinesin, the bacterial flagellar motor, and F1-ATPase.

Impact of fluorescent protein fusions on the bacterial flagellar motor.

Fluorescent fusion proteins open a direct and unique window onto protein function. However, they also introduce the risk of perturbation of the function of the native protein. Successful applications of fluorescent fusions therefore rely on a careful assessment and minimization of the side effects, but such insight is still lacking for many applications. This is particularly relevant in the study of the internal dynamics of motor proteins, where both the chemical and mechanical reaction coordinates can be affected. Fluorescent proteins fused to the stator of the Bacterial Flagellar Motor (BFM) have previously been used to unveil the motor subunit dynamics. Here we report the effects on single motors of three fluorescent proteins fused to the stators, all of which altered BFM behavior. The torque generated by individual stators was reduced while their stoichiometry remained unaffected. MotB fusions decreased the switching frequency and induced a novel bias-dependent asymmetry in the speed in the two directions. These effects could be mitigated by inserting a linker at the fusion point. These findings provide a quantitative account of the effects of fluorescent fusions to the stator on BFM dynamics and their alleviation- new insights that advance the use of fluorescent fusions to probe the dynamics of protein complexes.

Microresonator defects as sources of drifting cavity solitons.

Cavity solitons (CS) are localized structures appearing as single intensity peaks in the homogeneous background of the field emitted by a nonlinear (micro)resonator. In real devices, their position is strongly influenced by the presence of defects in the device structure. In this Letter we show that the interplay between these defects and a phase gradient in the driving field induces the spontaneous formation of a regular sequence of CSs moving in the gradient direction. Hence, defects behave as a device built-in CS source, where the CS generation rate can be set by controlling the system parameters.

Microscopy of extreme ultraviolet lithography masks with 13.2 nm tabletop laser illumination.

We report the demonstration of a reflection microscope that operates at 13.2 nm wavelength with a spatial resolution of 55+/-3 nm. The microscope uses illumination from a tabletop extreme ultraviolet laser to acquire aerial images of photolithography masks with a 20 s exposure time. The modulation transfer function of the optical system was characterized.

Highly coherent injection-seeded 13.2 nm tabletop soft x-ray laser.

We report a dramatic improvement of the spatial coherence and beam divergence (0.66 mrad) of a 13.2 nm wavelength Ni-like Cd tabletop laser by injection seeding the soft x-ray laser amplifier with high-harmonics pulses generated in a Ne gas jet. This phase coherent laser is an attractive light source for at-wavelength interferometry of extreme ultraviolet lithography optics and other applications.