The proton motive force determines Escherichia coli’s robustness to extracellular pH

Abstract

Maintaining intracellular homeostases is a hallmark of life, and key physiological variables, such as cytoplasmic pH, osmotic pressure, and proton motive force (PMF), are typically interdependent. Developing a mathematical model focused on these links, we predict that Escherichia coli uses proton-ion antiporters to generate an out-of-equilibrium plasma membrane potential and so maintain the PMF at the constant levels observed. The strength of the PMF consequently determines the range of extracellular pH over which the cell is able to preserve its near neutral cytoplasmic pH. In support, we concurrently measure the PMF and cytoplasmic pH in single cells and demonstrate both that decreasing the PMF’s strength impairs E. coli’s ability to maintain its pH and that artificially collapsing the PMF destroys the out-of-equilibrium plasma membrane potential. We further predict the observed ranges of extracellular pH for which three of E. coli’s antiporters are expressed, through defining their cost by the rate at which they divert imported protons from generating ATP. Taken together, our results suggest a new perspective on bacterial electrophysiology, where cells regulate the plasma membrane potential by changing the activities of antiporters to maintain both the PMF and cytoplasmic pH.