Segall J E, Ishihara A, Berg H C
J Bacteriol. 1985 Jan;161(1):51-9. doi: 10.1128/jb.161.1.51-59.1985.
Video techniques were used to record chemotactic responses of filamentous cells of Escherichia coli stimulated iontophoretically with aspartate. Long, nonseptate cells were produced from polyhook strains either by introducing a cell division mutation or by growth in the presence of cephalexin. Markers indicating rotation of flagellar motors were attached with anti-hook antibodies. Aspartate was applied by iontophoretic ejection from a micropipette, and the effects on the direction of rotation of the markers were measured. Motors near the pipette responded, whereas those sufficiently far away did not, even when the pipette was near the cell surface. The response of a given motor decreased as the pipette was moved away, but it did so less steeply when the pipette remained near the cell surface than when it was moved out into the external medium. This shows that there is an internal signal, but its range is short, only a few micrometers. These experiments rule out signaling by changes in membrane potential, by simple release or binding of a small molecule, or by diffusion of the receptor-attractant complex. A likely candidate for the signal is a protein or ligand that is activated by the receptor and inactivated as it diffuses through the cytoplasm. The range of the signal was found to be substantially longer in a cheZ mutant, suggesting that the product of the cheZ gene contributes to this inactivation.
利用视频技术记录经天冬氨酸离子电渗刺激的大肠杆菌丝状细胞的趋化反应。通过引入细胞分裂突变或在头孢氨苄存在下生长,从多钩菌株中产生长的、无隔膜的细胞。用抗钩抗体附着指示鞭毛马达旋转的标记物。通过微量移液器离子电渗喷射施加天冬氨酸,并测量其对标记物旋转方向的影响。靠近移液器的马达有反应,而那些距离足够远的马达则没有反应,即使移液器靠近细胞表面也是如此。随着移液器移开,给定马达的反应会减弱,但当移液器保持靠近细胞表面时,其减弱的程度比移到外部介质中时要小。这表明存在一种内部信号,但其范围很短,只有几微米。这些实验排除了通过膜电位变化、小分子的简单释放或结合以及受体 - 引诱剂复合物的扩散进行信号传导的可能性。信号的一个可能候选者是一种蛋白质或配体,它被受体激活并在通过细胞质扩散时失活。发现在cheZ突变体中信号的范围长得多,这表明cheZ基因的产物有助于这种失活。