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Nonlinear spectroscopy reveals how external pH controls permeation and accumulation of small molecules in two cellular compartments of Escherichia coli.

08:00 EDT 15th April 2019 | BioPortfolio

Summary of "Nonlinear spectroscopy reveals how external pH controls permeation and accumulation of small molecules in two cellular compartments of Escherichia coli."

Quantitative evaluation of small molecule permeation and accumulation in Gram-negative bacteria is important for drug development against them. While these measurements are commonly performed at physiological pH, Escherichia coli and many other Enterobacteriaceae infect human gastrointestinal and urinary tracts where they encounter different pH conditions. To understand how external pH affects permeation and accumulation of small molecules in E. coli cells, we apply second harmonic generation (SHG) spectroscopy using SHG-active antimicrobial compound malachite green as the probe molecule. Using SHG, we quantify periplasmic and cytoplasmic accumulations separately in live E. coli cells, which was never done before. Compartment-wise measurements reveal accumulation of the probe molecule in cytoplasm at physiological and alkaline pH, while entrapment in periplasm at weakly acidic pH and retention in external solution at highly acidic pH. Behind such disparity in localizations, up to two orders of magnitude reduction in permeability across the inner membrane at weakly acidic pH and outer membrane at highly acidic pH are found to play key roles. Our results unequivocally demonstrate the control of external pH over entry and compartment-wise distribution of small molecules in E. coli cells, which is a vital information and should be taken into account in antibiotic screening against E. coli and other Enterobacteriaceae members. In addition, our results demonstrate the ability of malachite green as an excellent SHG-indicator of changes of individual cell membrane and periplasm properties of live E. coli cells in response to external pH change from acidic to alkaline. This finding, too, has great importance as there is barely any other molecular probe which can provide similar information.

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This article was published in the following journal.

Name: Analytical chemistry
ISSN: 1520-6882
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