Apical uptake and transepithelial transport of sphingosine monomers through intact human intestinal epithelial cells: physicochemical and molecular modeling studies.
The mechanism of absorption of sphingosine was studied in human intestinal epithelial cells Caco-2 and HT-29-D4. The experiments were performed below the critical micellar concentration of sphingosine which was evaluated to 6 microM by surface tension measurements. [3H]Sphingosine uptake was not inhibited by Na+-free conditions, ATP depletion, L-cycloserine or methyl-beta-cyclodextrin, consistent with a passive diffusion mechanism independent of lipid raft integrity. Molecular modeling studies suggested that sphingosine can adopt two distinct conformations: a high-energy "snake-like" conformer in water and an extended low-energy conformer in lipid phases. We propose that the energy stored in the compressed snake-like conformer is transformed into kinetic energy, allowing: (i) the motion of sphingosine through the unstirred water layer bathing the mucosal enterocyte surface, and (ii) its insertion into the enterocyte brush border membrane. Dietary lipids that stabilized the extended sphingosine conformer in mixed micelles (e.g., cholesterol and sphingomyelin) induced a marked inhibition of sphingosine absorption.
Garmy
Nicolas
N
Laboratoire de Biochimie et Physicochimie des Membranes Biologiques, UMR-INRA 1111, Université Paul Cézanne, 13397 Marseille Cedex 20, France.
Taïeb
Nadira
N
Yahi
Nouara
N
Fantini
Jacques
J
D000255:Adenosine Triphosphate / Q000494:pharmacology; D001692:Biological Transport; D018938:Caco-2 Cells; D003523:Cycloserine / Q000494:pharmacology; D004058:Diffusion; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D019073:HT29 Cells; D006801:Humans; D007422:Intestines / Q000166:cytology*; D050356:Lipid Metabolism; D008855:Microscopy, Electron, Scanning; D008956:Models, Chemical; D012964:Sodium / Q000494:pharmacology; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism*; D013499:Surface Properties; D013500:Surface Tension; D047392:beta-Cyclodextrins / Q000494:pharmacology
D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D000255:Adenosine Triphosphate; D003523:Cycloserine; D012964:Sodium; D013110:Sphingosine