Na(+)-independent, H(+)-coupled transepithelial beta-alanine absorption by human intestinal Caco-2 cell monolayers.
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.
Thwaites
D T
DT
Department of Physiological Sciences, University of Newcastle upon Tyne Medical School, United Kingdom.
McEwan
G T
GT
Brown
C D
CD
Hirst
B H
BH
Simmons
N L
NL
D001692:Biological Transport; D002460:Cell Line; D004553:Electric Conductivity; D004848:Epithelium / Q000378:metabolism; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007408:Intestinal Absorption; D011522:Protons; D012964:Sodium / Q000494:pharmacology*; D015091:beta-Alanine / Q000378:metabolism*
D005452:Fluoresceins; D005456:Fluorescent Dyes; D011522:Protons; D015091:beta-Alanine; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D012964:Sodium