Zhao R, Amsler C D, Matsumura P, Khan S
Department of Physiology & Biophysics, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
J Bacteriol. 1996 Jan;178(1):258-65. doi: 10.1128/jb.178.1.258-265.1996.
Salmonella typhimurium FliG and FliM are two of three proteins known to be necessary for flagellar morphogenesis as well as energization and switching of flagellar rotation. We have determined FliG and FliM levels in cellular fractions and in extended flagellar basal bodies, using antibodies raised against the purified proteins. Both proteins were found predominantly in the detergent-solubilized particulate fraction containing flagellar structures. Basal flagellar fragments could be separated from partially constructed basal bodies by gel filtration chromatography. FliG and FliM were present in an approximately equimolar ration in all gel-filtered fractions. FliG and FliM copy numbers, estimated relative to that of the hook protein from the early fractions containing long, basal, flagellar fragments, were (means +/- standard errors) 41 +/- 10 and 37 +/- 13 per flagellum, respectively. Extended structures were present in the earliest identifiable basal bodies. Immunoelectron microscopy and immunoblot gel analysis suggested that the FliG and, to a less certain degree, the FliM contents of these structures were the same as those for the complete basal bodies. These facts are consistent with the postulate that FliG and FliM affect flagellar morphogenesis as part of the extended basal structure, formation of which is necessary for assembly of more-distal components of the flagellum. The determined stoichiometries will provide important constraints to modelling energization and switching of flagellar rotation.
鼠伤寒沙门氏菌的FliG和FliM是已知的鞭毛形态发生以及鞭毛旋转的供能和切换所必需的三种蛋白质中的两种。我们使用针对纯化蛋白产生的抗体,测定了细胞组分和延长的鞭毛基体中FliG和FliM的含量。两种蛋白质主要存在于含有鞭毛结构的去污剂可溶解的颗粒组分中。鞭毛基体片段可以通过凝胶过滤色谱法与部分构建的基体分离。在所有凝胶过滤组分中,FliG和FliM以大约等摩尔比存在。相对于来自含有长的、基体的、鞭毛片段的早期组分中的钩蛋白的拷贝数,估计的FliG和FliM拷贝数分别为每个鞭毛41±10和37±13(平均值±标准误差)。在最早可识别的基体中存在延长的结构。免疫电子显微镜和免疫印迹凝胶分析表明,这些结构中FliG的含量以及在较小程度上FliM的含量与完整基体中的相同。这些事实与以下假设一致:FliG和FliM作为延长的基体结构的一部分影响鞭毛形态发生,其形成对于鞭毛更远端组分的组装是必需的。所确定的化学计量将为鞭毛旋转的供能和切换建模提供重要的限制。