BACKGROUND & AIMS: Responses of the proton motive force (the driving force for protons) in Helicobacter pylori to varying medium pH may explain gastric colonization. The aim of this study was to determine the effect of external pH (pHout) on the proton motive force, the sum of the pH gradient, and the potential difference across the bacterial membrane. METHODS: Intracellular pH (pHin) was measured by bis-carboxyethyl-carboxy fluorescein fluorescence and transmembrane potential difference (PD) by fluorescent quenching of 3,3'-dipropyl thiadicarbocyanine iodide at differing pHout and was correlated with survival. RESULTS: PD was -131 +/- 0.36 mV (n = 3), and pHin was about 8.4 at loading pHout 7.0. PD increased as pHout was increased from 4.0 to 8.0, giving a constant proton motive force of about -220 mV. Outside these limits, PD collapsed irreversibly to zero. Addition of 5 mmol/L urea to weak buffer at pH 3.0 or 3.5 prevented irreversible collapse of PD by elevation of pHout caused by NH3 production. Urea addition to weak buffer at pH 7.0 collapsed the PD as urease activity increased the pHout to about 8.4. Survival was also limited to this range of pHout. CONCLUSIONS: H. pylori survives over the range of pHout where it maintains a proton motive force. The effect of urease activity on pHout, while allowing gastric survival in acidic media, may limit survival in nonacidic media.
BACKGROUND & AIMS: Responses of the proton motive force (the driving force for protons) in Helicobacter pylori to varying medium pH may explain gastric colonization. The aim of this study was to determine the effect of external pH (pHout) on the proton motive force, the sum of the pH gradient, and the potential difference across the bacterial membrane. METHODS: Intracellular pH (pHin) was measured by bis-carboxyethyl-carboxy fluorescein fluorescence and transmembrane potential difference (PD) by fluorescent quenching of 3,3'-dipropyl thiadicarbocyanine iodide at differing pHout and was correlated with survival. RESULTS: PD was -131 +/- 0.36 mV (n = 3), and pHin was about 8.4 at loading pHout 7.0. PD increased as pHout was increased from 4.0 to 8.0, giving a constant proton motive force of about -220 mV. Outside these limits, PD collapsed irreversibly to zero. Addition of 5 mmol/L urea to weak buffer at pH 3.0 or 3.5 prevented irreversible collapse of PD by elevation of pHout caused by NH3 production. Urea addition to weak buffer at pH 7.0 collapsed the PD as urease activity increased the pHout to about 8.4. Survival was also limited to this range of pHout. CONCLUSIONS:H. pylori survives over the range of pHout where it maintains a proton motive force. The effect of urease activity on pHout, while allowing gastric survival in acidic media, may limit survival in nonacidic media.
Authors: Yi Wen; David R Scott; Olga Vagin; Elmira Tokhtaeva; Elizabeth A Marcus; George Sachs Journal: J Bacteriol Date: 2018-06-25 Impact factor: 3.490