Trans-membrane Trafficking Unit, Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna, Kunigami, Okinawa, 904-0495, Japan.
Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, Washington, USA.
BMC Biol. 2017 Oct 27;15(1):97. doi: 10.1186/s12915-017-0438-7.
In a macro-molecular complex, any minor change may prove detrimental. For a supra-molecular nano-machine like the bacterial flagellum, which consists of several distinct parts with specific characteristics, stability is important. During the rotation of the bacterial flagellar motor, which is located in the membrane, the flagella rotate at speeds between 200 and 2000 rpm, depending on the bacterial species. The hook substructure of the bacterial flagellum acts as a universal joint connecting the motor to the flagellar filament. We investigated the formation of the bacterial flagellar hook and its overall stability between the FlgE subunits that make up the hook and attempted to understand how this stability differs between bacteria.
An intrinsically disordered segment plays an important role for overall hook stability and for its structural cohesion during motor rotation. The length of this linker segment depends on the species of bacteria; for Salmonella enterica and Campylobacter jejuni it is approximately 37 and 54 residues, respectively. Few residues of the linker are conserved and mutating the conserved residues of the linker yields non-flagellated cells. In the case of Campylobacter, which rotates its flagella at a speed much higher than that of Salmonella, shortening the linker leads to a rupture of the hook at its base, decreasing cell motility. Our experiments show that this segment is required for polymerization and stability of the hook, demonstrating a surprising role for a disordered region in one of the most finely tuned and closely studied macromolecular machines.
This study reveals a detailed functional characteristic of an intrinsically disordered segment in the hook protein. This segment evolved to fulfill a specific role in the formation of the hook, and it is at the core of the stability and flexibility of the hook. Its length is important in the case of bacteria with high-speed rotating flagella. Finding a way of disrupting this linker in Campylobacter might help in preventing infections.
在大分子复合物中,任何微小的变化都可能带来不利影响。对于像细菌鞭毛这样由几个具有特定特征的不同部分组成的超分子纳米机器来说,稳定性很重要。在位于膜中的细菌鞭毛马达旋转过程中,鞭毛的转速在 200 到 2000rpm 之间变化,具体取决于细菌种类。细菌鞭毛的钩状亚结构充当了将马达连接到鞭毛丝的万向节。我们研究了细菌鞭毛钩的形成及其组成钩的 FlgE 亚基之间的整体稳定性,并试图了解这种稳定性在不同细菌之间的差异。
一段固有无序的片段对于整体钩的稳定性及其在马达旋转过程中的结构内聚性起着重要作用。这个连接片段的长度取决于细菌的种类;对于肠炎沙门氏菌和空肠弯曲菌,其长度分别约为 37 和 54 个残基。连接片段的少数残基是保守的,突变连接片段的保守残基会导致鞭毛缺失。对于空肠弯曲菌,其鞭毛的旋转速度远高于肠炎沙门氏菌,缩短连接片段会导致钩在基部断裂,从而降低细胞的运动能力。我们的实验表明,这个片段对于钩的聚合和稳定性是必需的,这表明在最精细调节和最深入研究的大分子机器之一中,无序区域发挥了令人惊讶的作用。
本研究揭示了钩状蛋白中一段固有无序片段的详细功能特征。这个片段在钩的形成中进化出了特定的作用,是钩的稳定性和灵活性的核心。在具有高速旋转鞭毛的细菌中,其长度很重要。找到一种方法破坏空肠弯曲菌中的这个连接可能有助于预防感染。