Dataset Information

Essential ion binding residues for Na⁺ flow in stator complex of the Vibrio flagellar motor.

ABSTRACT: The bacterial flagellar motor is a unique supramolecular complex which converts ion flow into rotational force. Many biological devices mainly use two types of ions, proton and sodium ion. This is probably because of the fact that life originated in seawater, which is rich in protons and sodium ions. The polar flagellar motor in Vibrio is coupled with sodium ion and the energy converting unit of the motor is composed of two membrane proteins, PomA and PomB. It has been shown that the ion binding residue essential for ion transduction is the conserved aspartic acid residue (PomB-D24) in the PomB transmembrane region. To reveal the mechanism of ion selectivity, we identified essential residues, PomA-T158 and PomA-T186, other than PomB-D24, in the Na⁺-driven flagellar motor. It has been shown that the side chain of threonine contacts Na⁺ in Na⁺-coupled transporters. We monitored the Na⁺-binding specific structural changes using ATR-FTIR spectroscopy. The signals were abolished in PomA-T158A and -T186A, as well as in PomB-D24N. Molecular dynamics simulations further confirmed the strong binding of Na⁺ to D24 and showed that T158A and T186A hindered the Na⁺ binding and transportation. The data indicate that two threonine residues (PomA-T158 and PomA-T186), together with PomB-D24, are important for Na⁺ conduction in the Vibrio flagellar motor. The results contribute to clarify the mechanism of ion recognition and conversion of ion flow into mechanical force.

SUBMITTER: Onoue Y

PROVIDER: S-EPMC6677748 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Essential ion binding residues for Na<sup>+</sup> flow in stator complex of the Vibrio flagellar motor.

Onoue Yasuhiro Y Iwaki Masayo M Shinobu Ai A Nishihara Yasutaka Y Iwatsuki Hiroto H Terashima Hiroyuki H Kitao Akio A Kandori Hideki H Homma Michio M

Scientific reports 20190802 1

The bacterial flagellar motor is a unique supramolecular complex which converts ion flow into rotational force. Many biological devices mainly use two types of ions, proton and sodium ion. This is probably because of the fact that life originated in seawater, which is rich in protons and sodium ions. The polar flagellar motor in Vibrio is coupled with sodium ion and the energy converting unit of the motor is composed of two membrane proteins, PomA and PomB. It has been shown that the ion binding ...[more]

PMID: 31375690

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Project description:Bacterial flagella are the best-known rotational organelles in the biological world. The spiral-shaped flagellar filaments that extend from the cell surface rotate like a screw to create a propulsive force. At the base of the flagellar filament lies a protein motor that consists of a stator and a rotor embedded in the membrane. The stator is composed of two types of membrane subunits, PomA (similar to MotA in Escherichia coli) and PomB (similar to MotB in E. coli), which are energy converters that assemble around the rotor to couple rotation with the ion flow. Recently, stator structures, where two MotB molecules are inserted into the center of a ring made of five MotA molecules, were reported. This structure inspired a model in which the MotA ring rotates around the MotB dimer in response to ion influx. Here, we focus on the Vibrio PomB plug region, which is involved in flagellar motor activation. We investigated the plug region using site-directed photo-cross-linking and disulfide cross-linking experiments. Our results demonstrated that the plug interacts with the extracellular short loop region of PomA, which is located between transmembrane helices 3 and 4. Although the motor stopped rotating after cross-linking, its function recovered after treatment with a reducing reagent that disrupted the disulfide bond. Our results support the hypothesis, which has been inferred from the stator structure, that the plug region terminates the ion influx by blocking the rotation of the rotor as a spanner. IMPORTANCE The biological flagellar motor resembles a mechanical motor. It is composed of a stator and a rotor. The force is transmitted to the rotor by the gear-like stator movements. It has been proposed that the pentamer of MotA subunits revolves around the axis of the B subunit dimer in response to ion flow. The plug region of the B subunit regulates the ion flow. Here, we demonstrated that the ion flow was terminated by cross-linking the plug region of PomB with PomA. These findings support the rotation hypothesis and explain the role of the plug region in blocking the rotation of the stator unit.

Dataset Information

Essential ion binding residues for Na⁺ flow in stator complex of the Vibrio flagellar motor.

Publications

Essential ion binding residues for Na<sup>+</sup> flow in stator complex of the Vibrio flagellar motor.

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Dataset Information

Essential ion binding residues for Na+ flow in stator complex of the Vibrio flagellar motor.

Publications

Essential ion binding residues for Na<sup>+</sup> flow in stator complex of the Vibrio flagellar motor.

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OmicsDI is part of the ELIXIR infrastructure

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Essential ion binding residues for Na⁺ flow in stator complex of the Vibrio flagellar motor.