Na(+)-independent, H(+)-coupled transepithelial beta-alanine absorption by human intestinal Caco-2 cell monolayers

D T Thwaites, Gordon T.A. McEwan, C D Brown, B H Hirst, N L Simmons

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72 Citations (Scopus)

Abstract

beta-Alanine transport across intact human intestinal epithelial (Caco-2) cell layers has been investigated. In Na(+)-containing solutions, net absorptive flux of beta-alanine from apical-to-basal surfaces is small or absent, despite Na(+)-dependent intracellular beta-alanine accumulation across both apical and basal surfaces. Upon apical acidification (apical pH 6.0, basal pH 7.5), beta-alanine absorptive flux and accumulation across the apical surface are increased. In Na(+)-free conditions, a significant absorptive flux of beta-alanine is observed, which is markedly stimulated upon apical acidification (pH 6.0). Cellular accumulation of beta-alanine across the apical but not basal surface is observed in Na(+)-free conditions, and this is increased by acidic (pH 6.0) solutions. Absorptive beta-alanine flux in Na(+)-free conditions with acidic apical solutions displays saturation kinetics and competitive inhibition by alanine and glycine, but not valine or serine. Addition of 20 mM beta-alanine to the apical solution of epithelial monolayers loaded with the pH indicator 2',7'-bis(2-carboxyethyl-5(6)-carboxyfluorescein) causes a marked decrement in intracellular pH. beta-Alanine transport is also electrogenic, a concentration-dependent increase in an inward short circuit current being observed in voltage-clamped epithelial monolayers. We conclude that a proton-dependent, but Na(+)-independent, amino acid transporter is expressed at the apical membrane of human intestinal Caco-2 cells, and we provide direct evidence for amino acid-stimulated proton influx across the apical membrane in this intact epithelial cell system.
Original languageEnglish
Pages (from-to)18438-41
Number of pages4
JournalThe Journal of Biological Chemistry
Volume268
Issue number25
Publication statusPublished - 5 Sept 1993

Keywords

  • Biological Transport
  • Cell Line
  • Electric Conductivity
  • Epithelium
  • Fluoresceins
  • Fluorescent Dyes
  • Humans
  • Hydrogen-Ion Concentration
  • Intestinal Absorption
  • Protons
  • Sodium
  • beta-Alanine

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