Mechanism Of Force Generation Research Articles

Microtubule-based force generation.

Microtubules are vital to many important cell processes, such as cell division, transport of cellular cargo, organelle positioning, and cell migration. Owing to their diverse functions, understanding microtubule function is an important part of cell biological research that can help in combating various diseases. For example, microtubules are an important target of chemotherapeutic drugs such as paclitaxel because of their pivotal role in cell division. Many functions of microtubules relate to the generation of mechanical forces. These forces are generally either a direct result of microtubule polymerization/depolymerization or generated by motor proteins that move processively along microtubules. In this review, we summarize recent efforts to quantify and model force generation by microtubules in the context of microtubule function. WIREs Nanomed Nanobiotechnol 2017, 9:e1428. doi: 10.1002/wnan.1428 For further resources related to this article, please visit the WIREs website.

Clathrin-coat disassembly illuminates the mechanisms of Hsp70 force generation.

Hsp70s use ATP hydrolysis to disrupt protein:protein associations or move macromolecules. One example is Hsc70-mediated disassembly of clathrin coats that form on vesicles during endocytosis. We exploit the exceptional features of these coats to test three models—Brownian ratchet, power-stroke and entropic pulling—proposed to explain how Hsp70s transform their substrates. Our data rule out the ratchet and power-stroke models, and instead support a collision pressure mechanism whereby collisions between clathrin coat walls and Hsc70s drive coats apart. Collision pressure is the complement to the pulling force described in the entropic pulling model. We also find that self-association can augment collision pressure to allow disassembly of clathrin lattices predicted to resist disassembly. These results illuminate how Hsp70s generate the forces that transform their substrates.

Actin filaments growing against a barrier with fluctuating shape.

We study force generation by a set of parallel actin filaments growing against a nonrigid obstacle, in the presence of an external load. The filaments polymerize by either moving the whole obstacle, with a large energy cost, or by causing local distortion in its shape which costs much less energy. The nonrigid obstacle also has local thermal fluctuations due to which its shape can change with time and we describe this using fluctuations in the height profile of a one-dimensional interface with Kardar-Parisi-Zhang dynamics. We find the shape fluctuations of the barrier strongly affect the force generation mechanism. The qualitative nature of the force-velocity curve is crucially determined by the relative time scale of filament and barrier dynamics. The height profile of the barrier also shows interesting variation with the external load. Our analytical calculations within mean-field theory show reasonable agreement with our simulation results.

An unstructured mesh arbitrary Lagrangian-Eulerian unsteady incompressible flow solver and its application to insect flight aerodynamics

In this paper, an unstructured mesh Arbitrary Lagrangian-Eulerian (ALE) incompressible flow solver is developed to investigate the aerodynamics of insect hovering flight. The proposed finite-volume ALE Navier-Stokes solver is based on the artificial compressibility method (ACM) with a high-resolution method of characteristics-based scheme on unstructured grids. The present ALE model is validated and assessed through flow passing over an oscillating cylinder. Good agreements with experimental results and other numerical solutions are obtained, which demonstrates the accuracy and the capability of the present model. The lift generation mechanisms of 2D wing in hovering motion, including wake capture, delayed stall, rapid pitch, as well as clap and fling are then studied and illustrated using the current ALE model. Moreover, the optimized angular amplitude in symmetry model, 45°, is firstly reported in details using averaged lift and the energy power method. Besides, the lift generation of complete cyclic clap and fling motion, which is simulated by few researchers using the ALE method due to large deformation, is studied and clarified for the first time. The present ALE model is found to be a useful tool to investigate lift force generation mechanism for insect wing flight.

마이크로 머시닝에서의 공구 정렬과 가공정밀도

A micro end mill is one of the precise tools used in machining ultra-precision products such as microchannel and micropatterned mold. To achieve the required precision of these products, several studies investigated the cutting force, burr formation, and burr generation mechanism of micro end mills; however, there are few studies on the alignment of micro tools, which is the foundation of machining. Hence, in this investigation, relation expressions were derived to determine the relation between the misalignment parameters and the machining accuracy. At the same time, the effect of the machining parameters was analyzed using a multiple linear regression analysis and the analysis of variance. The results indicate that the tilting angle of a micro tool has more influence on the machining accuracy than other parameters.

Mesomechanical model and analysis of an artificial muscle functioning: role of Poisson’s ratio

The mechanism of force generation in a polymer monofilament actuator element with auxetic characteristics is modeled to assess the development and the optimization of a controlled drive based on the use of electrostrictive polymers. The monofilament is considered as a viscoelastic rod. By assuming a ‘sliding thread’ deformation occurring within the system, the variation of the monofilament length during the uniform contraction and force generated during a uniaxial mode of actuation have been obtained. The distribution of the axial stress was determined along the length of the monofilament at various stages during the uniform contraction. The rate of contraction reaches a maximum, together with a minimum of the stress intensity when the equivalent Poisson’s ratio of the actuator is negative.

Speech biomechanics: What have we learned and modeled since Joseph Perkell’s tongue model In 1974?

With his “physiologically oriented, dynamic model of the tongue, Joseph Perkell introduced in 1974 a new methodological approach to understanding the “relationships among phonetic models and the properties and capabilities of the speech-production mechanism.” This approach has guided a large part of our studies in the two last decades. In order to investigate how mechanical properties of the orofacial motor system constrain the degrees of freedom of speech articulation and contribute to shaping the speech signals exchanged between speakers and listeners, we, among other research groups, have developed increasingly more realistic 2D, and then 3D, finite element(FE) biomechanical models of the human vocal tract and face. After summarizing some of our modeling and simulation results that shed light on some basic characteristics of speech production, we present recent developments which aim to improve the realism of the models: evaluation of the links between the FE mesh structure (based either on tetrahedra, hexahedra, or mixed elements) and simulation accuracy; development of an active 3D element that simulates muscle mechanics and muscle force generation mechanisms; use of Diffusion Tensor Imaging to investigate muscle anatomy; design of an Atlas-based method (i.e., without manual image segmentation) for the automatic generation of subject-specific models.

Entropic forces drive contraction of cytoskeletal networks.

The cytoskeleton is a network of interconnected protein filaments, which provide a three-dimensional scaffold for cells. Remodeling of the cytoskeleton is important for key cellular processes, such as cell motility, division, or morphogenesis. This remodeling is traditionally considered to be driven exclusively by processes consuming chemical energy, such as the dynamics of the filaments or the action of molecular motors. Here, we review two mechanisms of cytoskeletal network remodeling that are independent of the consumption of chemical energy. In both cases directed motion of overlapping filaments is driven by entropic forces, which arise from harnessing thermal energy present in solution. Entropic forces are induced either by macromolecular crowding agents or by diffusible crosslinkers confined to the regions where filaments overlap. Both mechanisms increase filament overlap length and lead to the contraction of filament networks. These force-generating mechanisms, together with the chemical energy-dependent mechanisms, need to be considered for the comprehensive quantitative picture of the remodeling of cytoskeletal networks in cells.

Generation of mechanical force by grafted polyelectrolytes in an electric field.

We study theoretically and by means of molecular dynamics (MD) simulations the generation of mechanical force by grafted polyelectrolytes in an external electric field, which favors its adsorption on the grafting plane. The force arises in deformable bodies linked to the free end of the chain. Varying the field, one controls the length of the nonadsorbed part of the chain and hence the deformation of the target body, i.e., the arising force too. We consider target bodies with a linear force-deformation relation and with a Hertzian one. While the first relation models a coiled Gaussian chain, the second one describes the force response of a squeezed colloidal particle. The theoretical dependences of generated force and compression of the target body on an applied field agree very well with the results of MD simulations. The analyzed phenomenon may play an important role in future nanomachinery, e.g., it may be used to design nanovices to fix nanosized objects.

Mechanism of Cooperative Force Generations between Skeletal Myosins

To understand the molecular mechanism of cooperative force generation between skeletal myosin molecules, we measured forces generated by myosin-rod cofilaments, in which approximately 17 myosin molecules interact with single actins at the mixing ratio used in this study. Combined with results from the computational model, three factors are important for synchronization of power strokes between myosin motors. First, strain-dependent kinetics are necessary to couple mechanochemical cycles between myosins. Second, multiple power stroke states further enhance a chance of power stroke synchronization. Finally, the physiological ATP concentration is another important factor to enhance a chance of power stroke synchronization, since the strain-dependent transitions accompanied by the first or second power stroke are the rate limiting steps at higher ATP concentrations. Consequently, our computational model predicts that most of steps were generated by synchronous executions of power strokes between several myosin motors at 1 mM ATP, while they are generated primarily by single myosins. Thus, ensemble average curves of steps obtained from our model were distinctively different between 1 mM and 10 μM ATP. These results were consistent with experimental results, supporting our conclusions.

Super-Resolution Imaging of Slowly Depolymerizing Microtubules Reveals No Curved Protofilaments with a Lifetime Longer than Seconds

Microtubules are 25 nm diameter, micrometer long, cylindrical protein polymers involved in cell division, cell organization, and intracellular transport among other roles. Microtubule dynamics, that is, how microtubules grow and shrink (polymerization, catastrophe, depolymerization, and rescue) plays a role in each of these functions. Microtubule depolymerization (shrinkage), in particular, is proposed to involve a structural transition from straight protofilaments during microtubule polymerization to curled protofilaments at microtubule plus-ends during shrinkage. Direct evidence of this curved structure (sometimes called ‘ram's horns’) comes from static electron microscopy images and from unresolved intensity increases in light microscopy. As of now, however, no dynamic images of depolymerizing microtubules have directly resolved this ram's horns structure.We use STORM super-resolution microscopy on a custom-built microscope (the Warwick Open Source Microscope, WOSM) to image microtubules, in vitro, with a spatial resolution better than 20 nm and a temporal resolution of 10 seconds. We slow microtubule depolymerization to a similar timescale using sub-stoichiometric concentrations of paclitaxel or kinesin binding in rigor. Movies of these slowly depolymerizing microtubules show no curved protofilaments at microtubule plus ends. Instead, depolymerizing microtubules remain straight objects with a diameter of 25 nm. Our interpretation of these negative results are that the ram's horns structures have a lifetime shorter than seconds. An alternative hypothesis is that partial stabilization of microtubules is concomitant with a straightening of protofilaments at the shrinking end. This alternative suggests that proposed coupling mechanisms for force generation by depolymerizing microtubules must accommodate straight, rather than curved, protofilaments.

Quantitative nanoscale imaging of orientational order in biological filaments by polarized superresolution microscopy

Essential cellular functions as diverse as genome maintenance and tissue morphogenesis rely on the dynamic organization of filamentous assemblies. For example, the precise structural organization of DNA filaments has profound consequences on all DNA-mediated processes including gene expression, whereas control over the precise spatial arrangement of cytoskeletal protein filaments is key for mechanical force generation driving animal tissue morphogenesis. Polarized fluorescence is currently used to extract structural organization of fluorescently labeled biological filaments by determining the orientation of fluorescent labels, however with a strong drawback: polarized fluorescence imaging is indeed spatially limited by optical diffraction, and is thus unable to discriminate between the intrinsic orientational mobility of the fluorophore labels and the real structural disorder of the labeled biomolecules. Here, we demonstrate that quantitative single-molecule polarized detection in biological filament assemblies allows not only to correct for the rotational flexibility of the label but also to image orientational order of filaments at the nanoscale using superresolution capabilities. The method is based on polarized direct stochastic optical reconstruction microscopy, using dedicated optical scheme and image analysis to determine both molecular localization and orientation with high precision. We apply this method to double-stranded DNA in vitro and microtubules and actin stress fibers in whole cells.

Actin puts the squeeze on Drosophila glue secretion.

An actin filament coat promotes cargo expulsion from large exocytosing vesicles, but the mechanisms of coat formation and force generation have been poorly characterized. Elegant imaging studies of the Drosophila melanogaster salivary gland now reveal how actin and myosin are recruited, and show that myosin II forms a contractile 'cage' that facilitates exocytosis.

Evaluation of force generation mechanisms in natural, passive hydraulic actuators.

Pine cones are well known natural actuators that can move their scales upon humidity gradient. The mechanism manifests itself through a displacement easily observable by the naked eye, but coupled with stress generation. In ancient Egypt, wooden wedges were used to break soft blocks of stone by the generated swelling stress. The purpose of the present study is to evaluate the ability of pine cone scales to generate forces while being wetted. In our experiments, a blocking force of around 3N is measured depending on the position on the pine cone where the scales are extracted. A fairly good agreement is obtained when theoretical results based on bimetallic strip systems are compared with experimental data, even if overestimation is observed arising from the input data considered for dry tissues. Inspired by a simplified pine cone microstructure, a biocomposite analogue is manufactured and tested. Although an adequate blocking force can be generated, it has a lower value compared to natural pine cones which benefit from optimized swelling tissue content and interfacial bond strength between them. This study provides new insights to understand the generation of force by pine cones as well as to develop novel biocomposite functionalities.

Mechanism of Side Force Generation and Its Alleviation over a Slender Body

Experiments and computational studies were made on a slender body having a slender ratio of 16 at a freestream velocity of and Reynolds number of based on the base diameter so as to obtain the details of the flowfield that leads to the generation of side force at large angles of attack. It is observed that the large side force experienced at a high angle of attack of 50 deg is due to the presence of asymmetric vortices, which change its direction in the downstream. The vortices lift off from the surface due to the adverse pressure gradient. The major contribution of side force is mainly from an initial length limited to six times the base diameter, suggesting that efforts to reduce the side force by introducing any control method would be effective if used in the initial part of the body. Use of a circular trip having 5% height of the base diameter placed at a distance of 3.5 times the base diameter modifies the initial vortices and hence the local side force distribution over the body, which is mainly responsible for substantial reduction in the side force.

Fundamental Characteristics of AAA+ Protein Family Structure and Function.

Many complex cellular events depend on multiprotein complexes known as molecular machines to efficiently couple the energy derived from adenosine triphosphate hydrolysis to the generation of mechanical force. Members of the AAA+ ATPase superfamily (ATPases Associated with various cellular Activities) are critical components of many molecular machines. AAA+ proteins are defined by conserved modules that precisely position the active site elements of two adjacent subunits to catalyze ATP hydrolysis. In many cases, AAA+ proteins form a ring structure that translocates a polymeric substrate through the central channel using specialized loops that project into the central channel. We discuss the major features of AAA+ protein structure and function with an emphasis on pivotal aspects elucidated with archaeal proteins.

Hybrid Multi-Physics Modeling of an Ultra-Fast Electro-Mechanical Actuator

The challenges of an HVDC breaker are to generate impulsive forces in the order of hundreds of kilonewtons within fractions of a millisecond, to withstand the arising internal mechanical stresses and to transmit these forces via an electrically-insulating device to the contact system with minimum time delay. In this work, several models were developed with different levels of complexity, computation time and accuracy. Experiments were done with two mushroom-shaped armatures to validate the developed simulation models. It was concluded that although the electromagnetic force generation mechanism is highly sensitive to the mechanical response of the system, the developed first order hybrid model is able to predict the performance of the breaker with good accuracy.

Small Crowders Slow Down Kinesin-1 Stepping by Hindering Motor Domain Diffusion.

The dimeric motor protein kinesin-1 moves processively along microtubules against forces of up to 7 pN. However, the mechanism of force generation is still debated. Here, we point to the crucial importance of diffusion of the tethered motor domain for the stepping of kinesin-1: small crowders stop the motor at a viscosity of 5 mPa·s-corresponding to a hydrodynamic load in the sub-fN (~10^{-4} pN) range-whereas large crowders have no impact even at viscosities above 100 mPa·s. This indicates that the scale-dependent, effective viscosity experienced by the tethered motor domain is a key factor determining kinesin's functionality. Our results emphasize the role of diffusion in the kinesin-1 stepping mechanism and the general importance of the viscosity scaling paradigm in nanomechanics.

Structural Change in the Dynein Stalk Region Associated with Two Different Affinities for the Microtubule

Dynein is a large microtubule-based motor complex that requires tight coupling of intra-molecular ATP hydrolysis with the generation of mechanical force and track-binding activity. However, the microtubule-binding domain is structurally separated by about 15nm from the nucleotide-binding sites by a coiled-coil stalk. Thus, long-range two-way communication is necessary for coordination between the catalytic cycle of ATP hydrolysis and dynein's track-binding affinities. To investigate the structural changes that occur in the dynein stalk region to produce two different microtubule affinities, here we improve the resolution limit of the previously reported structure of the entire stalk region and we investigate structural changes in the dynein stalk and strut/buttress regions by comparing currently available X-ray structures. In the light of recent crystal structures, the basis of the transition from the low-affinity to the high-affinity coiled-coil registry is discussed. A concerted movement model previously reported by Carter and Vale is modified more specifically, and we proposed it as the open zipper model.

Microstructure actuation and gas sensing by the Knudsen thermal force

The generation of forces and moments on structures immersed in rarefied non-isothermal gas flows has received limited practical implementation since first being discovered over a century ago. The formation of significant thermal stresses requires both large thermal gradients and characteristic dimensions which are comparable to the gas molecular mean free path. For macroscopic geometries, this necessitates impractically high temperatures and very low pressures. At the microscale, however, these conditions are easily achieved, allowing the effects to be exploited, namely, for gas-property sensing and microstructure actuation. In this letter, we introduce and experimentally evaluate performance of a microelectromechanical in-plane Knudsen radiometric actuator, a self-contained device having Knudsen thermal force generation, sensing, and tuning mechanisms integrated onto the same platform. Sensitivity to ambient pressure, temperature gradient, as well as gas composition is demonstrated. Results are presented in terms of a non-dimensional force coefficient, allowing measurements to be directly compared to the previous experimental and computational data on out-of-plane cantilevered configurations.