Measurements of the Ion Channel Activity of the Transmembrane Stator Complex in the Bacterial Flagellar Motor

Methods Mol Biol. 2023:2646:83-94. doi: 10.1007/978-1-0716-3060-0_8.

Abstract

The bacterial flagellum is driven by a rotational motor located at the base of the flagellum. The stator unit complex conducts cations such as protons (H+) and sodium ions (Na+) along the electrochemical potential across the cytoplasmic membrane and interacts with the rotor to generate the rotational force. Escherichia coli and Salmonella have the H+-type stator complex, which serves as a transmembrane H+ channel that couples H+ flow through an ion channel to torque generation whereas Vibrio and some Bacillus species have the Na+-type stator complex. In this chapter, we describe how to measure the ion conductivity of the transmembrane stator complex over-expressed in E. coli cells using fluorescent indicators. Intensity measurements of fluorescent indicators using either a fluorescence spectrophotometer or microscope allow quantitative detection of changes in the intracellular ion concentrations due to the ion channel activity of the transmembrane protein complex.

Keywords: Bacterial flagellar motor; Fluorescence microscopy; Fluorescent protein; Ion channel; Ion conductivity; Spectroscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Flagella / metabolism
  • Ion Channels / metabolism
  • Ions / metabolism
  • Molecular Motor Proteins / metabolism
  • Protons
  • Vibrio alginolyticus* / metabolism

Substances

  • Protons
  • Ion Channels
  • Ions
  • Bacterial Proteins
  • Molecular Motor Proteins