Biological Molecular Motors Research Articles

Bidirectional Molecular Motors by Controlling Threading and Dethreading Pathways of a Linked Rotaxane.

Artificial molecular motors have been presented as models for biological molecular motors. In contrast to the conventional artificial molecular motors that rely on covalent bond rotation, molecular motors with mechanically interlocked molecules (MIM) have attracted considerable attention owing to their ability to generate significant rotational motion by dynamically shuttling macrocyclic components. The topology of MIM-type rotational molecular motors is currently limited to catenane structures, which require intricate synthetic procedures that typically produce a low synthetic yield. In this study, we develop a novel class of MIM-type molecular motors with a rotaxane-type topology. The switching of the threading/dethreading pathways of the linked rotaxane by protecting/deprotecting the bulky stopper group and changing the solvent polarity enables a net unidirectional rotation of the molecular motor. The threading/dethreading reaction rates were quantitatively evaluated through detailed spectroscopic investigations. Repeated net unidirectional rotation and switching of the direction of rotation were also achieved. Our findings demonstrate that linked rotaxanes can serve as MIM-type molecular motors with reversible rotational direction controlled by threading/dethreading reactions. These motors hold potential as components of molecular machinery.

Controlling Chaos: Periodic Defect Braiding in Active Nematics Confined to a Cardioid.

This work examines self-mixing in active nematics, a class of fluids in which mobile topological defects drive chaotic flows in a system comprised of biological filaments and molecular motors. We present experiments that demonstrate how geometrical confinement can influence the braiding dynamics of the defects. Notably, we show that confinement in cardioid-shaped wells leads to realization of the golden braid, a maximally efficient mixing state of exactly three defects with no defect creation or annihilation. We characterize the golden braid state using different measures of topological entropy and the Lyapunov exponent. In particular, topological entropy measured from the stretching rate of material lines agrees well with an analytical computation from braid theory. Increasing the size of the confining cardioid produces a transition from the golden braid, to the fully chaotic active turbulent state.

Design of artificial molecular motor inheriting directionality and scalability

Realizing artificial molecular motors with autonomous functionality and high performance is a major challenge in biophysics. Such motors not only provide new perspectives in biotechnology but also offer a novel approach for the bottom-up elucidation of biological molecular motors. Directionality and scalability are critical factors for practical applications. However, the simultaneous realization of both remains challenging. In this study, we propose a novel design for a rotary motor that can be fabricated using a currently available technology, DNA origami, and validate its functionality through simulations with practical parameters. We demonstrate that the motor rotates unidirectionally and processively in the direction defined by structural asymmetry, which induces kinetic asymmetry in motor motion. The motor also exhibits scalability, such that increasing the number of connections between the motor and stator allows for a larger speed, run length, and stall force.

Biosensing using antibody-modulated motility of actin filaments on myosin-coated surfaces

Motor proteins, such as myosin and kinesin, are biological molecular motors involved in force generation and intracellular transport within living cells. The characteristics of molecular motors, i.e., their motility over long distances, their capacity of transporting cargoes, and their very efficient energy consumption, recommend them as potential operational elements of a new class of dynamic nano-devices, with potential applications in biosensing, analyte concentrators, and biocomputation. A possible design of a biosensor based on protein molecular motor comprises a surface with immobilized motors propelling cytoskeletal filaments, which are decorated with antibodies, presented as side-branches. Upon biomolecular recognition of these branches by secondary antibodies, the ‘extensions’ on the cytoskeletal filaments can achieve considerable lengths (longer than several diameters of the cytoskeletal filament carrier), thus geometrically impairing or halting motility. Because the filaments are several micrometers long, this sensing mechanism converts an event in the nanometer range, i.e., antibody-antigen sizes, into an event in the micrometer range: the visualization of the halting of motility of microns-long cytoskeletal filaments. Here we demonstrate the proof of concept of a sensing system comprising heavy-mero-myosin immobilized on surfaces propelling actin filaments decorated with actin antibodies, whose movement is halted upon the recognition with secondary anti-actin antibodies. Because antibodies to the actin-myosin system are involved in several rare diseases, the first possible application for such a device may be their prognosis and diagnosis. The results also provide insights into guidelines for designing highly sensitive and very fast biosensors powered by motor proteins.

Macroscale Collagen‐Actomyosin Hybrid Actuator Built from Bioderived Materials

AbstractBiological muscles mainly consist of motor proteins that play essential roles and have a hierarchical structure covered with collagen that can transmit the tension to bones via tendons. The artificial reconstruction of motor proteins is highly expected from the viewpoint of engineering and biology. Although artificial muscles have been developed via synthetic and biological molecular motors, there is a considerable gap between the components of artificial and biological muscles, resulting in a significant obstacle to further scale‐up and force transfer through macroscale structures. To overcome these obstacles via bioderived biomolecules, this study proposes a collagen‐actomyosin hybrid soft actuator inspired by the structure of biological muscles. Collagen gel entrapping actomyosin can play roles in maintaining the actuator's shape and transmitting the tensile force. The generated forces reach the micro‐newton range, enabling the actuation of millimeter‐scale mechanical components. These properties may be helpful for the fabrication of soft robotic systems with advanced functionalities.

Unidirectional rotation of micromotors on water powered by pH-controlled disassembly of chiral molecular crystals

Biological and synthetic molecular motors, fueled by various physical and chemical means, can perform asymmetric linear and rotary motions that are inherently related to their asymmetric shapes. Here, we describe silver-organic micro-complexes of random shapes that exhibit macroscopic unidirectional rotation on water surface through the asymmetric release of cinchonine or cinchonidine chiral molecules from their crystallites asymmetrically adsorbed on the complex surfaces. Computational modeling indicates that the motor rotation is driven by a pH-controlled asymmetric jet-like Coulombic ejection of chiral molecules upon their protonation in water. The motor is capable of towing very large cargo, and its rotation can be accelerated by adding reducing agents to the water.

Optical Tweezers to Force Information out of Biological and Synthetic Systems One Molecule at a Time

Over the last few decades, in vitro single-molecule manipulation techniques have enabled the use of force and displacement as controlled variables in biochemistry. Measuring the effect of mechanical force on the real-time kinetics of a biological process gives us access to the rates, equilibrium constants and free-energy landscapes of the mechanical steps of the reaction; this information is not accessible by ensemble assays. Optical tweezers are the current method of choice in single-molecule manipulation due to their versatility, high force and spatial and temporal resolutions. The aim of this review is to describe the contributions of our lab in the single-molecule manipulation field. We present here several optical tweezers assays refined in our laboratory to probe the dynamics and mechano-chemical properties of biological molecular motors and synthetic molecular devices at the single-molecule level.

A new class of biological ion-driven rotary molecular motors with 5:2 symmetry.

Several new structures of three types of protein complexes, obtained by cryo-electron microscopy (cryo-EM) and published between 2019 and 2021, identify a new family of natural molecular wheels, the “5:2 rotary motors.” These span the cytoplasmic membranes of bacteria, and their rotation is driven by ion flow into the cell. They consist of a pentameric wheel encircling a dimeric axle within the cytoplasmic membrane of both Gram-positive and gram-negative bacteria. The axles extend into the periplasm, and the wheels extend into the cytoplasm. Rotation of these wheels has never been observed directly; it is inferred from the symmetry of the complexes and from the roles they play within the larger systems that they are known to power. In particular, the new structure of the stator complex of the Bacterial Flagellar Motor, MotA5B2, is consistent with a “wheels within wheels” model of the motor. Other 5:2 rotary motors are believed to share the core rotary function and mechanism, driven by ion-motive force at the cytoplasmic membrane. Their structures diverge in their periplasmic and cytoplasmic parts, reflecting the variety of roles that they perform. This review focuses on the structures of 5:2 rotary motors and their proposed mechanisms and functions. We also discuss molecular rotation in general and its relation to the rotational symmetry of molecular complexes.

The role of extended range of interactions in the dynamics of interacting molecular motors

Motor proteins, also known as biological molecular motors, play important roles in various intracellular processes. Experimental investigations suggest that molecular motors interact with each other during the cellular transport, but the nature of such interactions remains not well understood. Stimulated by these observations, we present a theoretical study aimed to understand the effect of the range of interactions on dynamics of interacting molecular motors. For this purpose, we develop a new version of the totally asymmetric simple exclusion processes in which nearest-neighbor as well as the next nearest-neighbor interactions are taken into account in a thermodynamically consistent way. A theoretical framework based on a cluster mean-field approximation, which partially takes correlations into account, is developed to evaluate the stationary properties of the system. It is found that fundamental current–density relations in the system strongly depend on the strength and the sign of interactions, as well as on the range of interactions. For repulsive interactions stronger than some critical value, a mean-field theoretical approach predicts that increasing the range of interactions might lead to a change from unimodal to trimodal dependence in the flux-density fundamental diagram. However, it is not fully supported by extensive Monte Carlo computer simulations that test theoretical predictions. Although in most ranges of parameters a reasonable agreement between theoretical calculations and computer simulations is observed, there are situations when the cluster mean-field approach fails to describe properly the dynamics in the system. Theoretical arguments to explain these observations are presented. Our theoretical analysis clarifies the microscopic picture of how the range of interactions influences the dynamics of interacting molecular motors.

Effects of defective motors on the active transport in biosensors powered by biomolecular motors

Motor proteins, such as myosin and kinesin, are biological molecular motors involved in force generation and intracellular transport in living cells. They were proposed to drive molecular shuttles for the active transport of analytes, thus significantly accelerating the sensing process of biosensors. Integrating motor proteins into biosensors requires their immobilisation on the operating surfaces. However, this process makes some motor proteins defective, slowing analyte detection. Here, we investigated the movements of molecular shuttles on surfaces in the presence of active and defective motors using a Brownian dynamics simulation, and elucidated the effects of defective motor proteins on the transport efficiency of the shuttles. We found that the motility of shuttles depends on the fraction of active motors relative to defective ones and that over 90% of the surface-bound motor proteins must remain active for efficient transport. The high fraction of active motors required for efficient transport can be attributed to the difference in the binding lifetimes of active and defective motors to shuttles. These results provide insights into how motors accumulate on sensor surfaces and set a guideline for the choice of polymer materials for biosensors powered by motor proteins.

A nanoscale reciprocating rotary mechanism with coordinated mobility control

Biological molecular motors transform chemical energy into mechanical work by coupling cyclic catalytic reactions to large-scale structural transitions. Mechanical deformation can be surprisingly efficient in realizing such coupling, as demonstrated by the F1FO ATP synthase. Here, we describe a synthetic molecular mechanism that transforms a rotary motion of an asymmetric camshaft into reciprocating large-scale transitions in a surrounding stator orchestrated by mechanical deformation. We design the mechanism using DNA origami, characterize its structure via cryo-electron microscopy, and examine its dynamic behavior using single-particle fluorescence microscopy and molecular dynamics simulations. While the camshaft can rotate inside the stator by diffusion, the stator’s mechanics makes the camshaft pause at preferred orientations. By changing the stator’s mechanical stiffness, we accelerate or suppress the Brownian rotation, demonstrating an allosteric coupling between the camshaft and the stator. Our mechanism provides a framework for manufacturing artificial nanomachines that function because of coordinated movements of their components.

The effect of local reversible dissociation of particles in interactive driven diffusive system

Motor proteins or biological molecular motors belong to a class of active enzymatic molecules that are responsible for transport and force generation in living cells. They typically operate in large teams and individual protein molecules interact with each other while moving along linear cytoskeleton filaments. Moreover, during their transportation the motors can reversibly dissociate from their tracks. Motivated by these observations, we propose a one dimensional totally asymmetric simple exclusion model for interacting particles that are allowed to reversibly dissociate/associate from a particular site far away from the system boundaries. A theoretical analysis of the model is based on cluster mean-field approximation that allows for a comprehensive description of the stationary properties in the system. It is found that the topology and nature of stationary phase diagrams for varying association/dissociation rates strongly depend on the sign and strength of interactions. Extensive Monte Carlo simulations are implemented to test our theoretical predictions.

Steady-State Dynamics of Exclusion Process with Local Reversible Association of Particles

Many biological processes are supported by special molecules, called motor proteins or molecular motors, that transport cellular cargoes along linear protein filaments and can reversibly associate to their tracks. Stimulated by these observations, we developed a theoretical model for collective dynamics of biological molecular motors that accounts for local association/dissociation events. In our approach, the particles interacting only via exclusion move along a lattice in the preferred direction, while the reversible associations are allowed at the specific site far away from the boundaries. Considering the association/dissociation site as a local defect, the inhomogeneous system is approximated as two coupled homogeneous sub-lattices. This allows us to obtain a full description of stationary dynamics in the system. It is found that the number and nature of steady-state phases strongly depend on the values of association and dissociation transition rates. Microscopic arguments to explain these observations, as well as biological implications, are also discussed. Theoretical predictions agree well with extensive Monte Carlo computer simulations.

Physical requirements for scaling up network-based biocomputation

The high energy consumption of electronic data processors, together with physical challenges limiting their further improvement, has triggered intensive interest in alternative computation paradigms. Here we focus on network-based biocomputation (NBC), a massively parallel approach where computational problems are encoded in planar networks implemented with nanoscale channels. These networks are explored by biological agents, such as biological molecular motor systems and bacteria, benefitting from their energy efficiency and availability in large numbers. We analyse and define the fundamental requirements that need to be fulfilled to scale up NBC computers to become a viable technology that can solve large NP-complete problem instances faster or with less energy consumption than electronic computers. Our work can serve as a guide for further efforts to contribute to elements of future NBC devices, and as the theoretical basis for a detailed NBC roadmap.

Optimal Rectification without Forward-Current Suppression by Biological Molecular Motor.

We experimentally show that biological molecular motor F_{1}-ATPase (F_{1}) implements an optimal rectification mechanism. The rectification mechanism hardly suppresses the synthesis of adenosine triphosphate by F_{1}, which is F_{1}'s physiological role, while inhibiting the unfavorable hydrolysis of adenosine triphosphate. This optimal rectification contrasts highly with that of a simple ratchet model, where the inhibition of the backward current is inevitably accompanied by the suppression of the forward current. Our detailed analysis of single-molecule trajectories demonstrates a novel but simple rectification mechanism of F_{1} with parallel landscapes and asymmetric transition rates.

A printable active network actuator built from an engineered biomolecular motor.

Leveraging the motion and force of individual molecular motors in a controlled manner to perform macroscopic tasks can provide substantial benefits to many applications, including robotics. Nonetheless, although millimetre-scale movement has been demonstrated with synthetic and biological molecular motors, their efficient integration into engineered systems that perform macroscopic tasks remains challenging. Here, we describe an active network capable of macroscopic actuation that is hierarchically assembled from an engineered kinesin, a biomolecular motor, and microtubules, resembling the contractile units in muscles. These contracting materials can be formed in desired areas using patterned ultraviolet illumination, allowing their incorporation into mechanically engineered systems, being also compatible with printing technologies. Due to the designed filamentous assembly of kinesins, the generated forces reach the micronewton range, enabling actuation of millimetre-scale mechanical components. These properties may be useful for the fabrication of soft robotic systems with advanced functionalities.

Effect of local dissociations in bidirectional transport of driven particles

Motivated by the complex processes of cellular transport when different types of biological molecular motors can move in opposite directions along protein filaments while also detaching from them, we developed a theoretical model of the bidirectional motion of driven particles. It utilizes a totally asymmetric simple exclusion process framework to analyze the dynamics of particles moving in opposite directions along the lattice of discrete sites while the particles might also dissociate from the filament in the bulk of the system. Mean-field theoretical arguments supported by extensive Monte Carlo simulations are presented in order to understand how the localized particle dissociations affect the bidirectional dynamics and spontaneous symmetry-breaking phenomena. It is found that changes in the amplitudes and in the symmetry of dissociation rates lead to significant modifications in the dynamic properties and in the stationary phase diagrams. These changes are explained using simple physical arguments. Our theoretical method clarifies some aspects of microscopic mechanisms of complex transport phenomena in biological systems.

Analysis of interactions in totally asymmetric exclusion process with site-dependent hopping rates: theory and simulations

Biological molecular motors are special enzymes that support biological processes such as intracellular transport, vesicle locomotion, RNA translation and many more. Experimental works suggest that the motor proteins interact among each other and moreover they experience a push by other motors during the intracellular transport. To incorporate these dynamics, we consider a variant of open one-dimensional totally asymmetric simple exclusion process with site-dependent hopping rates and interactions. The qualitative properties of our system do not depend on the hopping rate function. We utilize the simple mean-field, the cluster mean-field, the correlated cluster mean-field to theoretically calculate the stationary phase diagrams, density and the maximal particle current. The limitations of all the three theories are extensively discussed. It has been found that although the interactions do not change the number of phases in a phase diagram, it significantly changes the density profiles, the phase transition lines and the maximal particle current. The theoretical results obtained are supported by Monte Carlo simulations.

The role of dynamic defects in transport of interacting molecular motors

Motor proteins or biological molecular motors are special enzyme molecules that drive biological transport in living cells by moving cellular cargoes along linear protein filaments. The experimental evidences suggest that while performing their mechanical work biological molecular motors interact with each other, and there are other biological molecules on their tracks that influence their progression. Stimulated by these observations, we propose a one-dimensional totally asymmetric simple exclusion process with nearest-neighbor interactions and a dynamic defect that is allowed to reversibly bind and unbind at a specific site far away from the boundaries. A theoretical framework based on cluster mean-field approximation is adopted to determine the stationary properties of the system. The role of interactions and the effect the reversible defect associations on the dynamics of the system is discussed. It is found that three or less stationary phases can exist in the system, depending on the interaction strength, and only one of them is strongly affected by the defect association/dissociation dynamics. The theoretical results are validated through extensive Monte Carlo simulations.

Efficiencies of molecular motors: a comprehensible overview.

Many biological molecular motors can operate specifically and robustly at the highly fluctuating nano-scale. How these molecules achieve such remarkable functions is an intriguing question that requires various notions and quantifications of efficiency associated with the operations and energy transduction of these nano-machines. Here we give a short review of some important concepts of motor efficiencies, including the thermodynamic, Stokes, and generalized and transport efficiencies, as well as some implications provided by the thermodynamic uncertainty relations recently developed in nonequilibrium physics.