Energy source of flagellar type III secretion

Bacterial flagella contain a specialized secretion apparatus that functions to deliver the protein subunits that form the filament and other structures to outside the membrane. This apparatus is related to the injectisome used by many gram-negative pathogens and symbionts to transfer effector proteins into host cells; in both systems this export mechanism is termed 'type III' secretion. The flagellar secretion apparatus comprises a membrane-embedded complex of about five proteins, and soluble factors, which include export-dedicated chaperones and an ATPase, FliI, that was thought to provide the energy for export. Here we show that flagellar secretion in Salmonella enterica requires the proton motive force (PMF) and does not require ATP hydrolysis by FliI. The export of several flagellar export substrates was prevented by treatment with the protonophore CCCP, with no accompanying decrease in cellular ATP levels. Weak swarming motility and rare flagella were observed in a mutant deleted for FliI and for the non-flagellar type-III secretion ATPases InvJ and SsaN. These findings show that the flagellar secretion apparatus functions as a proton-driven protein exporter and that ATP hydrolysis is not essential for type III secretion.     

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.     

磁流变液铁磁颗粒的动力学分析

分析磁流变液铁磁颗粒的受力和运动特点对研究磁流变液微观机理至关重要。基于磁偶极子理论,比较全面地分析了磁流变液中的铁磁颗粒在外加磁场作用下所受到的力,并对这些力进行了比较。同时,研究了颗粒可能的运动方式。分析结果表明,磁力、载液的粘性阻力和颗粒间的排斥力是颗粒的主要受力,平动是颗粒的主要运动方式。并通过数值模拟手段,得到磁流变液在磁场作用下的链柱状微结构,从而证明该动力学分析的有效性。     

Active sliding between cytoplasmic microtubules

Microtubules are versatile cellular polymers that play a role in cell shape determination and mediate various motile processes such as ciliary and flagellar bending, chromosome movements and organelle transport. That a sliding microtubule mechanism can generate force has been demonstrated in highly ordered structures such as axonemes, and microtubule-based force generation almost certainly contributes to the function of mitotic and meiotic spindles. Most cytoplasmic microtubule arrays, however, do not exhibit the structural regularity of axonemes and some spindles, and often appear disorganized. Yet many cellular activities (such as shape changes during morphogenesis, axonal extension and spindle assembly) involve highly coordinated microtubule behaviour and possibly require force generated by an intermicrotubule sliding mechanism, or perhaps use sliding to move microtubules rapidly into a protrusion for stabilization. Here we show that active sliding between cytoplasmic microtubules can occur in microtubule bundles of the amoeba Reticulomyxa. A force-producing mechanism of this sort could be used by this organism to facilitate the extension of cell processes and to generate the dynamic movements of the cytoplasmic network.     

反井钻机ZX315镐型镶齿滚刀有效钻压分析

反井钻机在金属和非金属矿山坚硬岩石地层中钻进井筒时,在因反井钻杆和硐室空间问题限制钻机能力的情况下,经常需要采用略高于有效破岩钻压进行钻进,而通过全尺寸滚刀破岩实验或单齿压入实验可以判断有效钻压.选用ZX315镐型镶齿滚刀和单轴抗压强度141.6 MPa的花岗岩,进行直线往复式滚刀破岩实验对.在25～150 kN不同钻压时,通过观察破岩面、称重破碎岩碴、记录实验数据,分析得到钻压与破岩体积关系曲线拐点,进而获得有效钻压为100 kN,根据钻压与滚动力关系曲线得到滚刀的滚动摩擦因数为0.203,通过对岩石单轴抗压强度和有效钻压的拟合,得到单轴抗压强度与有效钻压的关系公式,可计算得到有效扭矩.通过对不同岩石的有效破岩钻压分析,得到有效破岩钻压与岩石单轴抗压强度的关系,并推算得出ZX315镐型镶齿滚刀的有效钻进钻压近似为岩石单轴抗压强度的75.48％.     

基于广义逆矩阵的机器人力与位置混合控制

推导了具有运动约束的机器人的拉格朗日动力学方程,该方程中包含了广义约束力。广义约束力被表达为机器人系统状态变量和广义输入力的显函数,力信息由此获得,因此无须使用力传感器即可同时控制机器人的力和位置。利用动力学冗余度（广义输入力的数目大于广义约束力的数目）,设计了基于广义逆矩阵的力与位置混合控制器。控制仿真结果表明,该控制方法具有较高的精度和较好的稳定性。     

Polar and lateral flagellar motors of marine Vibrio are driven by different ion-motive forces.

Various species of marine Vibrio produce two distinct types of flagella, each adapted for a different type of motility. A single, sheathed polar flagellum is suited for swimming in liquid medium, and numerous unsheathed lateral flagella, which are produced only under viscous conditions, are suited for swarming over viscous surfaces. Both types of flagella are driven by reversible motors embedded in the cytoplasmic membrane. Here we report that the energy source for the polar flagellar motor of Vibrio parahaemolyticus is the sodium-motive force, whereas the lateral flagellar motors are driven by the proton-motive force. This is evidence that two distinct types of flagella powered by different energy sources are functionally active in one cell.     

刀翼式PDC钻头的侧向力平衡设计

聚晶金刚石(PDC)钻头的侧向不平衡力是造成钻头涡动的主要原因之一,而侧向不平衡力取决于钻头的布齿结构,通过合理的布齿结构设计,可有效地控制侧向不平衡力的大小。在对PDC钻头进行受力分析的基础上,建立了PDC钻头受力计算模型,提出了通过调整刀翼周向位置使钻头的侧向不平衡力达到最小的优化设计方法。利用该方法进行了实例计算,结果表明,该设计方法能将PDC钻头的侧向不平衡力控制在钻压的1%以内。     

Spontaneous recovery after experimental manipulation of the plane of beat in sperm flagella

It is generally accepted that the oscillatory beating characteristic of sperm flagella is the result of an ATP-induced sliding between the doublet microtubules of the flagellar axoneme, with these longitudinal forces being converted into a lateral bending moment by resistive components of the structure that limit the displacement. However, little is known about the mechanisms that regulate this sliding among the nine doublets of the cylindrical axoneme to produce the coordinated planar bending waves required for efficient sperm propulsion. We have investigated these mechanisms with a new procedure in which the sperm head is held in the tip of a vibrating micropipette. Data obtained by gradually rotating the plane of imposed vibration around the sperm axis indicate that the pattern of active sliding between the outer doublet tubules can rotate relative to the sperm head, and suggest that this active sliding is regulated in part by the central tubule complex.     

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.     

平面桁架内力求解的“奇怪”现象分析

在所有理论力学教材中,平面任意力系有三个独立的平衡方程,这是众所周知的理论.但在某种情况下,却会出现一些特殊的现象.如用截面法求平面桁架内力的题目中,就出现有人称为“奇怪”的现象,即明明是一平面任意力系,有3个未知力,列出的3个平衡方程也无误,但怎么求解也求解不出结果来,许多学生甚至部分老师对此感到疑惑不解.对此给出了笔者的观点与看法,这实际上涉及到了一个特殊的平面任意力系独立平衡方程的问题.     

动静叶干涉中背压扰动叶栅气动力的降阶模型

现有航空发动机气动力降阶模型的研究主要集中在叶片颤振方向,对动静叶干涉引起叶栅气动力变化的问题没有讨论。其中背压扰动是动静叶干涉影响上游叶栅气动力的重要因素。通过对背压扰动的傅里叶分解,分别计算分解背压扰动所得各谐波引起的叶栅气动力响应,由此建立了基于谐波平衡法的背压扰动叶栅气动力降阶模型。在此基础上,研究了各谐波幅值对气动力降阶模型精度的影响。算例的结果表明:提出的方法能非常好地描述动静叶干涉中背压对上游叶栅气动力的影响;根据幅值大小选择主谐波,可以在不大幅降低气动力降阶模型精度的情况下减少待定参数的个数;利用该方法可准确确定任意时刻动静叶干涉中背压扰动叶栅的气动力。     

Sub-piconewton force fluctuations of actomyosin in vitro.

A new system has been developed for measuring the forces produced by a small number (less than 5-150) of myosin molecules interacting with a single actin filament in vitro. The technique can resolve forces of less than a piconewton and has a time resolution in the submillisecond range. It can thus detect fluctuations of force caused by individual molecular interactions. From analysis of these force fluctuations, the coupling between the enzymatic ATPase activity of actomyosin and the resulting mechanical impulses can be elucidated.     

Distinct roles of the FliI ATPase and proton motive force in bacterial flagellar protein export

Translocation of many soluble proteins across cell membranes occurs in an ATPase-driven manner. For construction of the bacterial flagellum responsible for motility, most of the components are exported by the flagellar protein export apparatus. The FliI ATPase is required for this export, and its ATPase activity is regulated by FliH; however, it is unclear how the chemical energy derived from ATP hydrolysis is used for the export process. Here we report that flagellar proteins of Salmonella enterica serovar Typhimurium are exported even in the absence of FliI. A fliH fliI double null mutant was weakly motile. Certain mutations in FlhA or FlhB, which form the core of the export gate, substantially improved protein export and motility of the double null mutant. Furthermore, proton motive force was essential for the export process. These results suggest that the FliH-FliI complex facilitates only the initial entry of export substrates into the gate, with the energy of ATP hydrolysis being used to disassemble and release the FliH-FliI complex from the protein about to be exported. The rest of the successive unfolding/translocation process of the substrates is driven by proton motive force.     

浅海结构物波浪力计算及实验研究

应用椭圆余弦波理论、双曲波理论及孤立波理论对作用于浅海海洋结构物的波浪力进行了计算 ,并在造波水槽中进行了波浪力及波压力实验研究。运用实验结果对传统的计算方法进行了修正 ,提出了静水面以上波压力的修正公式。研究表明 ,应用椭圆余弦波理论和双曲波理论计算理论波压力并对其进行修正 ,再将积分上限取为瞬时波面 ,积分后得出的波浪力与实验结果很接近。     

经编机连杆机构动力分析

本文介绍经编机连杆机构的动力分析。文中计算了当机器运转时,由于各构件的惯性力所引起的铰接点的约束反力以及传递至机架的振动力和振动力矩。文章中采用了两种方法:第一种方法是对机构中每一构件写出三个动力学方程,把这些方程写成矩阵形式,从而解得铰链点的约束反力和驱动力偶矩,进而计算振动力和振动力矩;第二种方法是利用达兰贝尔原理和能量公式直接写出振动力、振动力矩和驱动力偶矩的表达式。文中还提供了一些数据资料。     

固井封固系统初始作用力及其影响

结合建井过程和水泥浆的水化硬化过程对封固系统初始作用力进行分析。结果表明:固井作业完成时封固系统受到初始作用力的作用,初始作用力包括井眼附近地层承担的钻开井眼后重新分布的地应力、套管内部流体对套管的静液柱压力、一界面处水泥环对套管的挤压力及其反作用力、二界面处水泥环对地层的挤压力及其反作用力,作用在一界面处的初始作用力形成的初始应力稍小于地层孔隙压力;作用在二界面处的初始作用力形成的初始应力与地层孔隙压力相等;一、二界面处的初始作用力是水泥浆水化直至形成水泥石的过程中逐渐传递地层孔隙压力的结果。现场测量数据验证了一、二界面处初始作用力的存在。封固系统初始作用力的变化会对封固系统产生重要影响。     

Microtubule-associated protein 1C from brain is a two-headed cytosolic dynein

Dynein, an ATPase, is the force-generating protein in cilia and flagella. It has long been speculated that cytoplasmic microtubules contain a related enzyme involved in cell division or in intracellular organelle transport. A 'cytoplasmic dynein' has been described in sea urchin eggs, but because the egg stockpiles precursors for both cytoplasmic and ciliary microtubules, the role of this enzyme in the cell has remained unresolved. We recently found that the microtubule-associated protein (MAP) 1C (ref. 6) from brain is a microtubule-activated ATPase that produces force in the direction corresponding to retrograde organelle transport in the cell. MAP 1C has several similar properties to ciliary and flagellar dynein. Here we show directly, using scanning transmission electron microscopy, that MAP 1C is structurally equivalent to the ciliary and flagellar enzyme and is the long-sought cytoplasmic analogue of this enzyme.     

一种壁面牵引移动式玻璃幕墙清洗机器人

提出了一种新颖的平面玻璃幕墙清洗机器人结构设计原则,据此设计了2个自身无行走机构并依靠壁面牵引移动的清洗机器人系统.机器人吸附在壁面上,在楼顶卷扬机的牵引作用下依靠自身重力沿壁面下滑,同时完成清洗作业,并通过双负压吸盘的交替吸附跨越突起的水平窗框障碍.为提高机器人的工作能力,进行了作业安全分析,并研究了吸盘吸附力的调整控制方法.现场试验表明机器人能有效完成幕墙清洗任务,具有机构简单、效率高、成本低、清洗效果好的特点.