This study has demonstrated the existence of an L-proline-dependent (Na independent) proton flux at the apical membrane level of the eel intestinal absorbing cells. Using isolated eel enterocytes and the pH-sensitive fluorescent dye 2', 7'-bis-(2-carboxyethyl)-5-(and-6)-carboxy-fluorescein, acetoxymethyl ester (BCECF), it was shown that a 20 mM concentration of the imino acid L-proline in the extracellular medium determined an intracellular acidification of similar to0.28 pH units. However, neither sucrose nor other amino acids were able to significantly acidify the resting intracellular pH. A hyperbolic relationship between extracellular proline concentration and intracellular proton accumulation was observed. Using both isolated brush-border and basolateral membrane vesicles, it was demonstrated that this proline-proton cotransport mechanism was located at the apical membrane level only. In addition, the existence of a coupling mechanism between proline and proton fluxes was demonstrated by the observation that, in brush-border membrane vesicles, the presence of a pH gradient (pH(in) > pH(out)) stimulated the uptake of L-proline.

An L-proline-dependent proton flux is located at the apical membrane level of the eel enterocytes.

MARSIGLIANTE, Santo;STORELLI, Carlo;VILELLA, Sebastiano
2000-01-01

Abstract

This study has demonstrated the existence of an L-proline-dependent (Na independent) proton flux at the apical membrane level of the eel intestinal absorbing cells. Using isolated eel enterocytes and the pH-sensitive fluorescent dye 2', 7'-bis-(2-carboxyethyl)-5-(and-6)-carboxy-fluorescein, acetoxymethyl ester (BCECF), it was shown that a 20 mM concentration of the imino acid L-proline in the extracellular medium determined an intracellular acidification of similar to0.28 pH units. However, neither sucrose nor other amino acids were able to significantly acidify the resting intracellular pH. A hyperbolic relationship between extracellular proline concentration and intracellular proton accumulation was observed. Using both isolated brush-border and basolateral membrane vesicles, it was demonstrated that this proline-proton cotransport mechanism was located at the apical membrane level only. In addition, the existence of a coupling mechanism between proline and proton fluxes was demonstrated by the observation that, in brush-border membrane vesicles, the presence of a pH gradient (pH(in) > pH(out)) stimulated the uptake of L-proline.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11587/300378
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