Na+-independent lysine transport in human intestinal Caco-2 cells.
The nature of transepithelial and cellular transport of the dibasic amino acid lysine in human intestinal epithelial Caco-2 cells has been characterized. Intracellular accumulation of lysine across both the apical and basolateral membranes consists of a Na+-independent, membrane potential-sensitive uptake. Na+-independent lysine uptake at the basolateral membrane exceeds that at the apical membrane. Lysine uptake consists of both saturable and nonsaturable components. Na+-independent lysine uptake at both membranes is inhibited by lysine, arginine, alanine, histidine, methionine, leucine, cystine, cysteine and homoserine. In contrast, proline and taurine are without inhibitory effects at both membranes. Fractional Na+-independent lysine efflux from preloaded epithelial layers is greater at the basolateral membrane and shows trans-stimulation across both epithelial borders by lysine, arginine, alanine, histidine, methionine, and leucine but not proline and taurine. Na+-independent lysine influx (10 micron) in the presence of 10 mm homoserine shows further concentration dependent inhibition by lysine. Taken together, these data are consistent with lysine transport being mediated by systems bo,+, y+ and a component of very low affinity (nonsaturable) at both membranes. The relative contribution to lysine uptake at each membrane surface (at 10 micron lysine), normalized to total apical uptake (100%), is apical bo,+ (47%), y+ (27%) and the nonsaturable component (26%), and basal bo,+ (446%), y+ (276%) and the nonsaturable component (20%). Northern analysis shows hybridization of Caco-2 poly(A)+RNA with a human rBAT cDNA probe.
Thwaites
D T
DT
Department of Physiological Sciences, University of Newcastle upon Tyne, Medical School, Newcastle upon Tyne NE2 4HH, UK.
Markovich
D
D
Murer
H
H
Simmons
N L
NL
D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids / Q000235:genetics; D000704:Analysis of Variance; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D015152:Blotting, Northern; D018938:Caco-2 Cells / Q000502:physiology*; D002352:Carrier Proteins / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D004848:Epithelium / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000235:genetics; D012334:RNA, Neoplasm / Q000302:isolation & purification; D012964:Sodium / Q000378:metabolism* / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D012334:RNA, Neoplasm; C499489:SLC7A9 protein, human; D012964:Sodium; D008239:Lysine