Complete atomic model of the bacterial flagellar filament by electron cryomicroscopy

The bacterial flagellar filament is a helical propeller for bacterial locomotion. It is a helical assembly of a single protein, flagellin, and its tubular structure is formed by 11 protofilaments in two distinct conformations, L- and R-type, for supercoiling. The X-ray crystal structure of a flagellin fragment lacking about 100 terminal residues revealed the protofilament structure, but the full filament structure is still essential for understanding the mechanism of supercoiling and polymerization. Here we report a complete atomic model of the R-type filament by electron cryomicroscopy. A density map obtained from image data up to 4 A resolution shows the feature of alpha-helical backbone and some large side chains. The atomic model built on the map reveals intricate molecular packing and an alpha-helical coiled coil formed by the terminal chains in the inner core of the filament, with its intersubunit hydrophobic interactions having an important role in stabilizing the filament.     

Structure of the C-terminal domain of FliG, a component of the rotor in the bacterial flagellar motor.

Many motile species of bacteria are propelled by flagella, which are rigid helical filaments turned by rotary motors in the cell membrane. The motors are powered by the transmembrane gradient of protons or sodium ions. Although bacterial flagella contain many proteins, only three-MotA, MotB and FliG-participate closely in torque generation. MotA and MotB are ion-conducting membrane proteins that form the stator of the motor. FliG is a component of the rotor, present in about 25 copies per flagellum. It is composed of an amino-terminal domain that functions in flagellar assembly and a carboxy-terminal domain (FliG-C) that functions specifically in motor rotation. Here we report the crystal structure of FliG-C from the hyperthermophilic eubacterium Thermotoga maritima. Charged residues that are important for function, and which interact with the stator protein MotA, cluster along a prominent ridge on FliG-C. On the basis of the disposition of these residues, we present a hypothesis for the orientation of FliG-C domains in the flagellar motor, and propose a structural model for the part of the rotor that interacts with the stator.     

Structure of the bacterial flagellar protofilament and implications for a switch for supercoiling

The bacterial flagellar filament is a helical propeller constructed from 11 protofilaments of a single protein, flagellin. The filament switches between left- and right-handed supercoiled forms when bacteria switch their swimming mode between running and tumbling. Supercoiling is produced by two different packing interactions of flagellin called L and R. In switching from L to R, the intersubunit distance ( approximately 52 A) along the protofilament decreases by 0.8 A. Changes in the number of L and R protofilaments govern supercoiling of the filament. Here we report the 2.0 A resolution crystal structure of a Salmonella flagellin fragment of relative molecular mass 41,300. The crystal contains pairs of antiparallel straight protofilaments with the R-type repeat. By simulated extension of the protofilament model, we have identified possible switch regions responsible for the bi-stable mechanical switch that generates the 0.8 A difference in repeat distance.     

Direct observation of steps in rotation of the bacterial flagellar motor

The bacterial flagellar motor is a rotary molecular machine that rotates the helical filaments that propel many species of swimming bacteria. The rotor is a set of rings up to 45 nm in diameter in the cytoplasmic membrane; the stator contains about ten torque-generating units anchored to the cell wall at the perimeter of the rotor. The free-energy source for the motor is an inward-directed electrochemical gradient of ions across the cytoplasmic membrane, the protonmotive force or sodium-motive force for H+-driven and Na+-driven motors, respectively. Here we demonstrate a stepping motion of a Na+-driven chimaeric flagellar motor in Escherichia coli at low sodium-motive force and with controlled expression of a small number of torque-generating units. We observe 26 steps per revolution, which is consistent with the periodicity of the ring of FliG protein, the proposed site of torque generation on the rotor. Backwards steps despite the absence of the flagellar switching protein CheY indicate a small change in free energy per step, similar to that of a single ion transit.     

汽车空间多万向节传动的动态优化设计

从汽车实际行驶的动态工况出发，提出了对汽车空间多万向节传动最佳布置的数学模型，选择了合适的优化算法，编制了计算程序．实例计算表明，降低由万向节传动的不等速性引起的传动系的振动和噪声，采用动态优化方法比采用静态优化方法效果更好     

作业型ROV推进器动力学建模与响应分析

为研究深海作业型遥控水下机器人（remotely operated vehicle,ROV）液压推进器控制系统的动力学响应特性,建立了一种考虑螺旋桨动态负载影响的伺服阀控制液压推进器动力学系统的数学模型,提出一种伺服阀控制液压推进器的马达流量、压力、扭矩、转速、螺旋桨转矩和推力的求解方法.通过数值仿真,分析了不同控制电压下伺服阀、液压马达和螺旋桨的动态响应过程及特点,建立了推力分配方法中推力简化约束模型,并得到了期望推力和推进器控制电压之间函数关系的数学模型.与推进器水池试验结果相比,本文仿真结果准确可信.这种完整和准确的液压推进器动力学系统的数学模型,对实际水下机器人和动力定位船舶的运动控制方法、推力分配策略及推进器控制的研究,具有一定的指导意义和工程价值.      

Abrupt changes in flagellar rotation observed by laser dark-field microscopy

Bacteria such as Escherichia coli and Salmonella typhimurium swim by rotating their flagella, each of which consists of an external helical filament and a rotary motor embedded in the cell surface. The function of the flagellar motor has been examined mainly by tethering the flagellar filament to a glass slide and observing the resultant rotation of the cell body. But under these conditions the motor operates at a very low speed (about 10 r.p.s.) owing to the unnaturally high load conditions inherent in this technique. Lowe et al. analysed the frequency of light scattered from swimming cells to estimate the average rotation speed of flagellar bundles of E. coli as about 270 r.p.s. To analyse motor function in more detail, however, measurement of high-speed rotation of a single flagellum (at low load) with a temporal resolution better than 1 ms is needed. We have now developed a new method--laser dark-field microscopy--which fulfils these requirements. We find that although the average rotation speed of S. typhimurium flagella is rather stable, there are occasional abrupt slowdowns, pauses and reversals (accomplished within 1 ms). These changes were frequently observed in mutants defective in one of the motor components (called the switch complex), suggesting that this component is important not only in switching rotational direction but also in torque generation or regulation.     

In situ structure of the complete Treponema primitia flagellar motor

The bacterial flagellar motor is an amazing nanomachine: built from approximately 25 different proteins, it uses an electrochemical ion gradient to drive rotation at speeds of up to 300 Hz (refs 1, 2). The flagellar motor consists of a fixed, membrane-embedded, torque-generating stator and a typically bidirectional, spinning rotor that changes direction in response to chemotactic signals. Most structural analyses so far have targeted the purified rotor, and hence little is known about the stator and its interactions. Here we show, using electron cryotomography of whole cells, the in situ structure of the complete flagellar motor from the spirochaete Treponema primitia at 7 nm resolution. Twenty individual motor particles were computationally extracted from the reconstructions, aligned and then averaged. The stator assembly, revealed for the first time, possessed 16-fold symmetry and was connected directly to the rotor, C ring and a novel P-ring-like structure. The unusually large size of the motor suggested mechanisms for increasing torque and supported models wherein critical interactions occur atop the C ring, where our data suggest that both the carboxy-terminal and middle domains of FliG are found.     

Unidirectional molecular motor on a gold surface

Molecules capable of mimicking the function of a wide range of mechanical devices have been fabricated, with motors that can induce mechanical movement attracting particular attention. Such molecular motors convert light or chemical energy into directional rotary or linear motion, and are usually prepared and operated in solution. But if they are to be used as nanomachines that can do useful work, it seems essential to construct systems that can function on a surface, like a recently reported linear artificial muscle. Surface-mounted rotors have been realized and limited directionality in their motion predicted. Here we demonstrate that a light-driven molecular motor capable of repetitive unidirectional rotation can be mounted on the surface of gold nanoparticles. The motor design uses a chiral helical alkene with an upper half that serves as a propeller and is connected through a carbon-carbon double bond (the rotation axis) to a lower half that serves as a stator. The stator carries two thiol-functionalized 'legs', which then bind the entire motor molecule to a gold surface. NMR spectroscopy reveals that two photo-induced cis-trans isomerizations of the central double bond, each followed by a thermal helix inversion to prevent reverse rotation, induce a full and unidirectional 360 degrees rotation of the propeller with respect to the surface-mounted lower half of the system.     

Stoichiometry and turnover in single, functioning membrane protein complexes

Many essential cellular processes are carried out by complex biological machines located in the cell membrane. The bacterial flagellar motor is a large membrane-spanning protein complex that functions as an ion-driven rotary motor to propel cells through liquid media. Within the motor, MotB is a component of the stator that couples ion flow to torque generation and anchors the stator to the cell wall. Here we have investigated the protein stoichiometry, dynamics and turnover of MotB with single-molecule precision in functioning bacterial flagellar motors in Escherichia coli. We monitored motor function by rotation of a tethered cell body, and simultaneously measured the number and dynamics of MotB molecules labelled with green fluorescent protein (GFP-MotB) in the motor by total internal reflection fluorescence microscopy. Counting fluorophores by the stepwise photobleaching of single GFP molecules showed that each motor contains approximately 22 copies of GFP-MotB, consistent with approximately 11 stators each containing two MotB molecules. We also observed a membrane pool of approximately 200 GFP-MotB molecules diffusing at approximately 0.008 microm2 s(-1). Fluorescence recovery after photobleaching and fluorescence loss in photobleaching showed turnover of GFP-MotB between the membrane pool and motor with a rate constant of the order of 0.04 s(-1): the dwell time of a given stator in the motor is only approximately 0.5 min. This is the first direct measurement of the number and rapid turnover of protein subunits within a functioning molecular machine.     

Adaptation at the output of the chemotaxis signalling pathway

In the bacterial chemotaxis network, receptor clusters process input, and flagellar motors generate output. Receptor and motor complexes are coupled by the diffusible protein CheY-P. Receptor output (the steady-state concentration of CheY-P) varies from cell to cell. However, the motor is ultrasensitive, with a narrow operating range of CheY-P concentrations. How the match between receptor output and motor input might be optimized is unclear. Here we show that the motor can shift its operating range by changing its composition. The number of FliM subunits in the C-ring increases in response to a decrement in the concentration of CheY-P, increasing motor sensitivity. This shift in sensitivity explains the slow partial adaptation observed in mutants that lack the receptor methyltransferase and methylesterase and why motors show signal-dependent FliM turnover. Adaptive remodelling is likely to be a common feature in the operation of many molecular machines.     

考虑吊钩组升降的伸缩臂起重机动力学模型研究

建立了一种模拟伸缩臂起重机转台回转,吊臂变幅、伸缩,吊钩升降的运动学、动力学模型.利用光滑连续的正余弦函数描述伸缩臂起重机的运动,使用拉格朗日方法列出吊钩组的动力学方程,并用时变约束方程描述吊钩钢丝绳的升降长度,最终组成微分代数方程,使用约束稳定化方法进行求解.通过算例验证了伸缩臂起重机动力学模型的正确性.该模型可以为伸缩臂起重机的复合动作控制研究及虚拟吊装方案设计提供参考.     

Torque-generating units of the flagellar motor of Escherichia coli have a high duty ratio

Rotation of the bacterial flagellar motor is driven by an ensemble of torque-generating units containing the proteins MotA and MotB. Here, by inducing expression of MotA in motA- cells under conditions of low viscous load, we show that the limiting speed of the motor is independent of the number of units: at vanishing load, one unit turns the motor as rapidly as many. This result indicates that each unit may remain attached to the rotor for most of its mechanochemical cycle, that is, that it has a high duty ratio. Thus, torque generators behave more like kinesin, the protein that moves vesicles along microtubules, than myosin, the protein that powers muscle. However, their translation rates, stepping frequencies and power outputs are much higher, being greater than 30 microm s(-1), 12 kHz and 1.5 x 10(5) pN nm s(-1), respectively.     

基于Matlab-Simulink的舰船综合电力推进系统仿真

为了优化舰船综合电力推进系统的性能,减少研制风险,基于M atlab-S im u link环境,建立了由十二相同步电机、电压型逆变器、螺旋桨及船舶负载组成的推进系统的各部分及系统的仿真模型。通过变压变频控制的十二相同步电动机带螺旋桨及船舶负载系统的仿真对模型进行了验证。为了提高舰船综合电力推进系统的性能,推导了十二相同步电动机直轴气隙磁链观测器的模型,籍此构建了十二相同步电动机矢量控制方案。仿真结果表明,各模型建立正确,十二相同步电动机矢量控制方案能够实现转矩与磁链控制解耦,能够满足舰船电力推进系统动态性能要求。     

球笼式等速万向节的接触应力分析及优化设计

以球笼式等速万向节作为研究对象,运用三维建模软件SolidWorks建立实体模型并进行简化,利用ANSYS对该万向节的两种工况做了有限元接触应力分析,找出其薄弱部位,通过改进滚道进行优化设计,缓解了应力集中现象,降低了最大应力,提高了万向节使用寿命。     

计算旋转螺旋面三维边界层的一种积分方法

本文将计算二维层流边界层的Thwaites方法和计算二维湍流边界层的Truckenbrodt方法推广,用以求解旋转系统的“准三维”边界层方程,得到了计算旋转螺旋面上三维边界层的一种可迭代求解的积分方法.应用此方法进行了两例计算.计算结果与理论精确解或实验测量的对比表明,本文方法是有效的.     

用升力面方法估算螺旋桨空泡

本文提出了用数值升力面理论求解三维空泡螺旋桨问题的准定常方法.表征螺旋桨负荷、厚度和空泡的奇点系及满足边界条件的控制点均置于拱弧面上。尾涡模型划分为过渡区和远尾流区,过渡区用圆锥螺旋面来模拟尾涡片的变形现象,远尾流区简化为桨叶数的集中等螺距梢涡和一根直线毂涡模型。空泡模型用半闭式.计算结果与实验值吻合良好。     

调距桨锁轴拖带工况最小拖桨阻力和水动力矩

采用计算流体力学方法,对某典型5叶调距桨锁轴工况拖桨阻力以及相应的水动力矩随螺距和进流速度的变化特性进行了数值计算,分析了这些特性的流体力学机理. 数值计算基于有限体积法,通过数值求解螺旋桨周围三维黏性不可压缩流场RANS方程来模拟螺旋桨在各种工况下的流动特性. 计算结果表明：来流速度相等时该调距桨在最大正车螺距时拖桨阻力最大,其幅值约占同航速下船体阻力的80%,水动力矩也最大;在最大倒车螺距时拖桨阻力最小,其幅值约占同航速下船体阻力的50%,水动力矩比最大正车螺距时显著减小;零螺距时拖桨阻力大小居中,而水动力矩最小,接近为零. 上述结论可为船舶动力装置部分桨工况时联控曲线的设计和锁轴机构的设计提供理论参考.     

基于ADAMS的焊接机器人轨迹规划

通过Pro/E建立六关节弧焊机器人的CAD模型,导入ADAMS软件并通过点驱动的方式,求解各关节角度随时间的变化;通过ADAMS的后处理模块,将测量所得各关节角度随时间的变化曲线转换为样条曲线,在各关节上添加驱动电机,观察求解的准确性;求解所得的各关节角度随时间变化的数据转换为相应的脉冲函数关系,在物理样机上进行驱动。     

轿车拖钩强度分析

文章采用有限元方法对拖钩强度进行分析以某轿车的拖钩为例,重点介绍了拖钩强度分析的具体流程,通过与目标值的对比,对其结构强度进行评价;并根据分析结果确定了优化拖钩结构的方案;对优化后的结构重新进行分析,仿真计算结果表明,拖钩的强度得到了提高.