Characteristics of light chains of Chara myosin revealed by immunological investigation

Chara myosin is plant myosin responsible for cytoplasmic streaming and moves actin filaments at 60 µm/s, which is the fastest of all myosins examined. The neck of the myosin molecule has usually mechanical and regulatory roles. The neck of Chara myosin is supposed to bind six light chains, but, at present, we have no knowledge about them. We found Ca⁺⁺-calmodulin activated Chara myosin motility and its actin-activated ATPase, and actually bound with the Chara myosin heavy chain, indicating calmodulin might be one of candidates for Chara myosin light chains. Antibody against essential light chain from Physarum myosin, and antibodies against Chara calmodulin and chicken myosin light chain from lens membranes reacted with 20 kDa and 18 kDa polypeptides of Chara myosin preparation, respectively. Correspondingly, column purified Chara myosin had light chains of 20 kDa, and 18 kDa with the molar ratio of 0.7 and 2.5 to the heavy chain, respectively.     

Transportation of drug–gold nanocomposites by actinomyosin motor system

Nanotechnology is playing an important role in drug delivery to overcome limitations of conventional drug delivery systems in terms of solubility, in vivo stability, pharmacokinetics, and bio-distribution. The controlled transportation of drug into the cell and within the cell is a major challenge to be addressed. Cellular molecular motors have been exploited for their cargo carrying capacity for various applications including engineering and health care. Combination of nanotechnology and biomolecular motors can address some of the challenges in drug delivery. In the present study, transportation of drug nanocomposites has been demonstrated. Nanocomposites of 6-mercaptopurine and levodopa drugs (cancer and Parkinson’s disease, respectively) were prepared with gold nanoparticles (GNPs) by covalent attachment and these nanocomposites were attached to actin filaments. These nanocomposites were in-turn transported by actin filaments on myosin tracks. Characterization of drug nanocomposites formation was done by UV–Vis spectroscopy, field emission scanning electron microscopy, transmission electron microscopy, and confocal microscopy. GNP composites of 6-mercaptopurine and levodopa were formed by sulfide and amide bond formation, respectively. Average velocity of actin filament attached to nanocomposites was found to be 3.17 and 3.89 μm/s for levodopa and 6-mercaptopurine, respectively, as compared to actin filaments with velocity of 4.0–6.0 μm/s. Three concepts have been proposed for the study of drug transportation into the cell based on polycationic complex formation, interaction of actin with cellular myosin and Biomolecular Adaptor for Retrograde Transport (BART) technology. The aspects of this study heads toward the development of an approach to utilize molecular motors for nanoscale transportation endogenously.     

Monte Carlo Simulation of Kinesin Movement with a Lattice Model

Kinesin is a processive double-headed molecular motor that moves along a microtubule by taking about 8nm steps. It generally hydrolyzes one ATP molecule for taking each forward step. The processive movement of the kinesin molecular motors is numerically simulated with a lattice model. The motors are considered as Brownian particles and the ATPase processes of both heads are taken into account. The Monte Carlo simulation results agree well with recent experimental observations, especially on the relation of velocity versus ATP and ADP concentrations.     

驱动蛋白单向运动机理

驱动蛋白马达在实验和理论上已被进行了广泛的研究. 然而, 其行进运动的微观机理仍未确定. 在本文中我们基于化学、力学和电学耦合提出了一个交臂模型来描述这种行进运动. 在该模型中，驱动蛋白两个头的ATP水解化学反应速率由作用在其颈上的力（包括内部弹性力和外部负荷）来调控. 在低外部负荷情况下, 驱动蛋白的后头的ATP水解化学反应速率远大于前头的速率, 因而两个头在ATP水解化学反应和力学周期循环中是协调的且马达以每步消耗一个ATP的方式的行走. 在大的前向负荷情况下, 两个头的ATP水解化学反应速率变得可比拟, 因而两个头在ATP水解化学反应和力学周期循环中不再很好地协调. 该模型与驱动蛋白的结构研究结果以及ATP水解化学反应路径一致. 利用此模型所估算的驱动力（约5.8 pN）与实验结果（5~7.5 pN）一致. 所估算的每步中的运动时间（约10）也与实验测量值（0~50）符合. 解释了已观察到的每步（8nm）分为两个半步的现象. 所得到的运动速度-负荷曲线与已有的实验结果一致.     

Stochastic force generation by small ensembles of myosin II motors

Forces in the actin cytoskeleton are generated by small groups of nonprocessive myosin II motors for which stochastic effects are highly relevant. Using a cross-bridge model with the assumptions of fast power-stroke kinetics and equal load sharing between equivalent states, we derive a one-step master equation for the activity of a finite-sized ensemble of mechanically coupled myosin II motors. For constant external load, this approach yields analytical results for duty ratio and force-velocity relation as a function of ensemble size. We find that stochastic effects cannot be neglected for ensemble sizes below 15. The one-step master equation can be used also for efficient computer simulations with linear elastic external load and reveals the sequence of buildup of force and ensemble rupture that is characteristic for reconstituted actomyosin contractility.     

Membrane waves driven by actin and Myosin

We present a model which couples the membrane with the protrusive forces of actin polymerization and contractile forces of molecular motors, such as myosin. The actin polymerization at the membrane is activated by freely diffusing membrane proteins that have a spontaneous curvature. Molecular motors are recruited to the polymerizing actin filaments, from a constant reservoir, and produce a contractile force. All the forces and variables are treated in the linear limit. Our results show that for convex membrane proteins the myosin activity gives rise to robust transverse membrane waves, similar to those observed on different cells.     

Experimental studies of the myosin-actin motor

The mechanical processes generated by the molecular motors, myosin and actin, were measured using single molecule imaging, manipulation, and nanometry techniques. It was shown that the mechanical events of myosin are not tightly coupled with the ATP hydrolysis reaction and that myosin molecules move stochastically. These results indicate that the movement of myosin is driven by thermal motion rather than structural changes occurring in the myosin molecules. Thermal Brownian motion must be biased using the energy released from the hydrolysis of ATP. Thus, the molecular motors can harness thermal energy to perform mechanical work efficiently. Received: 20 November 2001 / Accepted: 11 February 2002 / Published online: 22 April 2002     

Mechanochemical model for myosinⅡdimer that can explain the spontaneous oscillatory contraction of muscle

The spontaneous oscillatory contraction(SPOC) of myofibrils is the essential property inherent to the contractile system of muscle. Muscle contraction results from cyclic interactions between actin filament and myosin II which is a dimeric motor protein with two heads. Taking the two heads of myosin II as an indivisible element and considering the effects of cooperative behavior between the two heads on rate constants in the mechanochemical cycle, the present work proposes the tenstate mechanochemical cycle model for myosin II dimer. The simulations of this model show that the proportion of myosin II in different states periodically changes with time, which results in the sustained oscillations of contractive tension, and serves as the primary factor for SPOC. The good fit of this model to experimental results suggests that the cooperative interaction between the two heads of myosin II dimer may be one of the underlying mechanisms for muscle contraction.     

Double-temperature ratchet model and current reversal of coupled Brownian motors

On the basis of the transport features and experimental phenomena observed in studies of molecular motors, we propose a double-temperature ratchet model of coupled motors to reveal the dynamical mechanism of cooperative transport of motors with two heads, where the interactions and asynchrony between two motor heads are taken into account. We investigate the collective unidirectional transport of coupled system and find that the direction of motion can be reversed under certain conditions. Reverse motion can be achieved by modulating the coupling strength, coupling free length, and asymmetric coefficient of the periodic potential, which is understood in terms of the effective potential theory. The dependence of the directed current on various parameters is studied systematically. Directed transport of coupled Brownian motors can be manipulated and optimized by adjusting the pulsation period or the phase shift of the pulsation temperature.     

Covalent immobilization of myosin forin-vitro motility of actin

The present study reports the covalent immobilization of myosin on glass surface andin-vitro motility of actin-myosin biomolecular motor. Myosin was immobilized on poly-L-lysine coated glass using heterobifunctional cross linker EDC and characterized by AFM. Thein-vitro motility of actin was carried out on the immobilized myosin. It was observed that velocity of actin over myosin increases with increasing actin concentration (0.4–1.0 mg/ml) and was found in the range of 0.40–3.25 μm/s. The motility of actinmyosin motor on artificial surfaces is of immense importance for developing nanodevices for healthcare and engineering applications     

Motility States of Molecular Motors Engaged in a Stochastic Tug-of-War

Intracellular transport is mediated by molecular motors that pull cargos along cytoskeletal filaments. Many cargos move bidirectionally and are transported by two teams of motors which move into opposite directions along the filament. We have recently introduced a stochastic tug-of-war model for this situation. This model describes the motion of the cargo as a Markov process on a two-dimensional state space defined by the numbers of active plus and active minus motors. In spite of its simplicity, this tug-of-war model leads to a complex dependence of the cargo motility on the motor parameters. We present new numerical results for the dependence on the number of involved motors. In addition, we derive a simple and intuitive sharp maxima approximation, from which one obtains the cargo motility state from only four simple inequalities. This approach provides a fast and reliable method to determine the cargo motility.     

Transport properties of coupled Brownian particles confined to a microchannel in an asymmetric periodic potential

On the basis of the transport features and experimental phenomena observed in studies of molecular motors, we investigated an overdamped Brownian motion of two coupled particles with an asymmetric periodic potential in a two-dimensional microchannel theoretically and numerically, to reveal the dynamical mechanism of cooperative transport of particles with two heads, where the interactions between two particles are taken into consideration. Moreover, while moving in a confined structure, Brownian particles also could exhibit peculiar kinetic behavior. The dependence of directed current on various parameters is systematically studied. Our results indicate that the direction of motion can be reversed by modulating the coupling strength, free length, and microchannel width. In addition, we have achieved the conditions of forward motion in this study. That is, when the interparticle average horizontal interval Δx > 0.25L, where L is the spatial period of the external potential, the forward motion of coupled Brownian particles effected by the synchronized noise and confined to a microchannel can be generated in the strong-coupling case.     

向生命学习新的机制

根据有关分子马达的重要实验结果,明确指出分子马达的运动过程是一个化学、电学、力学3种过程互相耦合的生物学过程。主要介绍有关kinesin的力产生机制的研究现状以及在这方面的研究。On the basis of the important experimental results of molecular motors, it was polnted out that the moving process of molecular motors is a coupling biological process of chemical-electrical-mechanical processes. This clever mechanism of energy conversion on the molecular level with several processes coupled together had never been observed before. The understanding of this new mechanism is an important step towards the understanding of life and an important content of what we can learn from life. We introduced here the status of the investigations on the mechanism for the force generation of kinesin and the studies of the authors in this field     

Dynamic instability of collective myosin Ⅱ motors

Some kinds of muscles can oscillate spontaneously,which is related to the dynamic instability of the collective motors.Based on the two-state ratchet model and with consideration of the motor stiffness,the dynamics of collective myosin Ⅱmotors are studied.It is shown that when the motor stiffness is small,the velocity of the collective motors decreases monotonically with load increasing.When the motor stiffness becomes large,dynamic instability appears in the forcevelocity relationship of the collective-motor transport.For a large enough motor stiffness,the zero-velocity point lies in the unstable range of the force-velocity curve,and the motor system becomes unstable before the motion is stopped,so spontaneous oscillations can be generated if the system is elastically coupled to its environment via a spring.The oscillation frequency is related to the motor stiffness,motor binding rate,spring stiffness,and the width of the ATP excitation interval.For a medium motor stiffness,the zero-velocity point lies outside the unstable range of the force-velocity curve,and the motion will be stopped before the instability occurs.     

肌球蛋白Ⅵ分子马达周期势场下的弹性扩散模型

肌球蛋白Ⅵ分子马达因其特殊的结构及胞内功能,其动力学原理成为研究的热点. 从肌球蛋白Ⅵ自身结构和实验现象出发,建立其弹性扩散模型,并通过Monte Carlo方法分析了肌球蛋白Ⅵ满足朗之万方程的随机动力学行为. 结果表明,在环境噪声作用下,具有弹性势能和轨道周期势能的肌球蛋白Ⅵ可以进行梯跳运动和有效的输运,但负载力会减弱分子马达系统的输运能力;当弹性系数一定时,弹性链越长平均速度越小,当弹性链长度一定时,合理选择弹性系数平均速度可达到最大值;另外,负载力的存在使肌球蛋白Ⅵ在接触位点的平均驻留时间呈指数增加. 关键词： 分子马达 肌球蛋白Ⅵ 朗之万方程 弹性扩散模型     

Trapping and injecting single domain walls in magnetic wire by local fields

A single domain wall (DW) moves at linearly increasing velocity under an increasing homogeneous drive magnetic field. Present experiments show that the DW is braked and finally trapped at a given position when an additional antiparallel local magnetic field is applied. That position and its velocity are further controlled by suitable tuning of the local field. In turn, the parallel local field of small amplitude does not significantly affect the effective wall speed at long distance, although it generates tail-to-tail and head-to-head pairs of walls moving along opposite directions when that field is strong enough.     

基于蒙特卡罗方法的4H-SiC(0001)面聚并台阶形貌演化机理

针对SiC外延生长中微观原子动力学过程,建立了一个三维蒙特卡罗模型来研究偏向■或■方向4H-SiC(0001)邻晶面上台阶形貌演化过程,并且利用Burton-Cabera-Frank理论分析了其形成机理.在蒙特卡罗模型中,首先建立了一个计算4H-SiC晶体生长过程的晶格网格,用来确定Si原子和C原子晶格坐标以及联系它们之间的化学键;其次,考虑了原子在台阶面上的吸附、扩散,原子在台阶边上的附着、分离以及传输等过程;最后,为了更加详细地捕捉微观原子在晶体表面的动力学过程信息,该模型把Si原子和C原子分别对待,同时还考虑了能量势垒对吸附原子影响.模拟结果表明:在偏向■方向的4H-SiC(0001)邻晶面,有一个晶胞高度的聚并台阶形貌形成,而对于偏向■方向的邻晶面,出现了半个晶胞高度的聚并台阶形貌,该模拟结果与实验中观察到的结果相符合.最后,利用Burton-Cabera-Frank理论对聚并台阶形貌演化机理进行了讨论.     

Force generation by polymerizing microtubules

Polymerization ratchets formed by the assembly of actin filaments and microtubules are possibly the simplest realizations of biological thermal ratchets. A variety of experimental evidence exists that significant forces are generated by these processes, but quantitative studies lag far behind similar studies for molecular motors such as kinesin and myosin. Here we present a discussion of the theory of polymerization ratchets as well as experimental techniques used in our laboratory for the study of forces generated by single growing microtubules. Data obtained with these techniques provide us with valuable information that may eventually allow us to distinguish between different models for the growth of microtubules. Received: 15 January 2002 / Accepted: 11 February 2002 / Published online: 22 April 2002     

非水体系中金电极表面增强拉曼光谱研究新进展

利用共焦显微拉曼系统,进行了甲醇溶液中硫氰酸根离子（SCN^-）在金电极上吸附行为的表面增强拉曼光谱（SERS）研究,结果表明：SCN^-与电极之间的相互作用非常强烈,在整个研究电位范围内都可以检测到其SERS信号。在－0．1～－0．6V区间内,SCN^-主要是以S端吸附在电极表面：而在－0．7V～－1．2V区间内,SCN^-主要是以N端吸附在电极表面。