Effect of hydrogen ion-gradient on carrier-mediated transport of glycylglycine across brush border membrane vesicles from rabbit small intestine.

Glycylglycine (Gly-Gly) transport across rabbit small intestinal brush border membrane vesicles was studied in the presence and absence of a Na+ gradient or a pH gradient. The transport was found to be entirely independent of Na+ but significantly stimulated by lowering extravesicular pH (pHo). The maximum stimulation was seen at pHo 5.5, where the uptake rate was about 2-times higher than the control value. FCCP, a protonophore, abolished the stimulating effect of low pHo, and low pHo conditions without a pH gradient did not stimulate the uptake rate. Overshoot uptake of Gly-Gly was observed when a pH gradient of 2 pH units was imposed across the vesicular membrane. Valinomycin-induced inside-negative K+ diffusion potential also had a stimulating effect on the uptake, and fluorescence measurements of vesicular suspensions containing diS-C3-(5) revealed the occurrence of depolarization of the vesicular membranes when Gly-Gly was added to the suspensions. Kinetic study showed that a pH gradient caused a decrease of Kt for Gly-Gly without affecting Vmax. All the data obtained indicate that Gly-Gly transport is independent of Na+, dependent on a H+ gradient, and electrogenic, suggesting the mechanism of cotransport with H+.