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 (pH
o). The maximum stimulation was seen at pH
o 5.5, where the uptake rate was about 2-times higher than the control value. FCCP, a protonophore, abolished the stimulating effect of low pH
o, and low pH
o 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-C
3-(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 K
t for Gly-Gly without affecting V
max. 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
+.
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