Reid Stuart W, Leake Mark C, Chandler Jennifer H, Lo Chien-Jung, Armitage Judith P, Berry Richard M
Clarendon Laboratory, Department of Physics, University of Oxford, South Parks Road, Oxford OX1 3PU, United Kingdom.
Proc Natl Acad Sci U S A. 2006 May 23;103(21):8066-71. doi: 10.1073/pnas.0509932103. Epub 2006 May 12.
Torque is generated in the rotary motor at the base of the bacterial flagellum by ion translocating stator units anchored to the peptidoglycan cell wall. Stator units are composed of the proteins MotA and MotB in proton-driven motors, and they are composed of PomA and PomB in sodium-driven motors. Strains of Escherichia coli lacking functional stator proteins produce flagella that do not rotate, and induced expression of the missing proteins leads to restoration of motor rotation in discrete speed increments, a process known as "resurrection." Early work suggested a maximum of eight units. More recent indications that WT motors may contain more than eight units, based on recovery of disrupted motors, are inconclusive. Here we demonstrate conclusively that the maximum number of units in a motor is at least 11. Using back-focal-plane interferometry of 1-mum polystyrene beads attached to flagella, we observed at least 11 distinct speed increments during resurrection with three different combinations of stator proteins in E. coli. The average torques generated by a single unit and a fully induced motor were lower than previous estimates. Speed increments at high numbers of units are smaller than those at low numbers, indicating that not all units in a fully induced motor are equivalent.
扭矩由锚定在肽聚糖细胞壁上的离子转运定子单元在细菌鞭毛基部的旋转马达中产生。在质子驱动的马达中,定子单元由MotA和MotB蛋白组成,而在钠驱动的马达中,它们由PomA和PomB组成。缺乏功能性定子蛋白的大肠杆菌菌株产生不旋转的鞭毛,缺失蛋白的诱导表达会导致马达以离散的速度增量恢复旋转,这一过程称为“复活”。早期研究表明最多有八个单元。基于对受损马达的恢复情况,最近有迹象表明野生型马达可能包含超过八个单元,但尚无定论。在这里,我们确凿地证明马达中的单元最大数量至少为11个。通过对附着在鞭毛上的1微米聚苯乙烯珠进行背焦平面干涉测量,我们在大肠杆菌中使用三种不同的定子蛋白组合进行复活过程中观察到至少11个不同的速度增量。单个单元和完全诱导的马达产生的平均扭矩低于先前的估计值。单元数量较多时的速度增量小于数量较少时的速度增量,这表明完全诱导的马达中的并非所有单元都是等效的。