Biochemical characterization of the flagellar stator-associated inner membrane protein FliL from Vibrio alginolyticus
Biochemical characterization of the flagellar stator-associated inner membrane protein FliL from Vibrio alginolyticus
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DOI：
10.1093/jb/mvw076
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发表时间：
2017-04-01
影响因子：
2.7
通讯作者：
Homma, Michio
中科院分区：
文献类型：
--
作者：
Kumar, Ananthanarayanan;Isumi, Miyu;Homma, Michio
鞭毛马达嵌入在细胞包膜中，并在定子和转子之间的相互作用下旋转。旋转由通过定子的离子流提供动力。一种名为FliL的单跨膜蛋白与鞭毛马达中的扭矩产生相关。我们建立了溶藻弧菌FliL的大肠杆菌过表达系统，溶藻弧菌是一种具有钠驱动极鞭毛的海洋细菌。我们成功地表达，纯化和结晶的CA。17 kDa全长FliL蛋白，并产生仅表达ca. FliL的14 kDa周质区（Delta TM FliL）。生物化学表征和NMR分析表明，Delta TM FliL与自身弱相互作用形成寡聚体。我们推测，所观察到的动态相互作用可能参与了鞭毛运动功能的FliL的作用。
The flagellar motor is embedded in the cell envelope and rotates upon interaction between the stator and the rotor. The rotation is powered by ion flow through the stator. A single transmembrane protein named FliL is associated with torque generation in the flagellar motor. We established an Escherichia coli over-expression system for FliL of Vibrio alginolyticus, a marine bacterium that has a sodium-driven polar flagellum. We successfully expressed, purified, and crystallized the ca. 17 kDa full-length FliL protein and generated a construct that expresses only the ca. 14 kDa periplasmic region of FliL (Delta TM FliL). Biochemical characterization and NMR analysis revealed that Delta TM FliL weakly interacted with itself to form an oligomer. We speculate that the observed dynamic interaction may be involved in the role of FliL in flagellar motor function.