Transport of a tripeptide, Gly-Pro-Hyp, across the porcine intestinal brush-border membrane.
The transcellular transport of oligopeptides across intestinal epithelial cells has attracted considerable interest in investigations into how biologically active peptides express diverse physiological functions in the body. It has been postulated that the tripeptide, Gly-Pro-Hyp, which is frequently found in collagen sequences, exhibits bioactivity. However, the mechanism of uptake of dietary di- and tripeptides by intestinal epithelial cells is not well understood. In this study, we used porcine brush-border membrane (BBM) vesicles to assess Gly-Pro-Hyp uptake, because these vesicles can structurally and functionally mimic in vivo conditions of human intestinal apical membranes. The present study demonstrated the time-dependent degradation of this tripeptide into the free-form Gly and a dipeptide, Pro-Hyp, on the apical side of the BBM vesicles. In parallel with the hydrolysis of the tripeptide, the dipeptide Pro-Hyp was identified in the BBM intravesicular space environment. We found that the transcellular transport of Pro-Hyp across the BBM was inhibited by the addition of a competitive substrate (Gly-Pro) for peptide transporter (PEPT1) and was pH-dependent. These results indicate that Gly-Pro-Hyp can be partially hydrolyzed by the brush-border membrane-bound aminopeptidase N to remove Gly, and that the resulting Pro-Hyp is, in part, transported into the small intestinal epithelial cells via the H+-coupled PEPT1. Gly-Pro-Hyp cannot cross the epithelial apical membrane in an intact form, and Pro-Hyp is highly resistant to hydrolysis by intestinal mucosal apical proteases.
Aito-Inoue
Misako
M
Department of Food Sciences and Nutritional Health, Kyoto Prefectural University, 1-5 Nakaragi-cho, Shimogamo, Kyoto 606-8522, Japan.
Lackeyram
Dale
D
Fan
Ming Z
MZ
Sato
Kenji
K
Mine
Yoshinori
Y
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004847:Epithelial Cells / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D008871:Microvilli / Q000378:metabolism*; D008954:Models, Biological; D009842:Oligopeptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D052616:Solid Phase Extraction; D013552:Swine; D013997:Time Factors
D009842:Oligopeptides; C088188:glycyl-prolyl-hydroxyproline