Apical secretion and sialylation of soluble dipeptidyl peptidase IV are two related events.
The role of glycans in the apical targeting of proteins in epithelial cells remains a debated question. We have expressed the mouse soluble dipeptidyl peptidase IV (DPP IV ectodomain) in kidney (MDCK) and in intestinal (Caco-2) epithelial cell lines, as a model to study the role of glycosylation in apical targeting. The mouse DPP IV ectodomain was secreted mainly into the apical medium by MDCK cells. Exposure of MDCK cells to GalNac-alpha-O-benzyl, a drug previously described as an inhibitor of mucin O-glycosylation, produced a protein with a lower molecular weight. In addition this treatment resulted in a decreased apical secretion and an increased basolateral secretion of mouse DPP IV ectodomain. When expressed in Caco-2 cells, the mouse DPP IV ectodomain was secreted mainly into the basolateral medium. However, BGN was still able to decrease the amount of apically secreted protein and to increase its basolateral secretion. Neuraminidase digestion showed that the most striking effect of BGN was a blockade of DPP IV sialylation in both MDCK and Caco-2 cells. These results indicate that a specific glycosylation step, namely, sialylation, plays a key role in the control of the apical targeting of a secreted DPP IV both in MDCK and Caco-2 cells.
Slimane
T A
TA
INSERM U 538, CHU St. Antoine, Paris, France.
Lenoir
C
C
Sapin
C
C
Maurice
M
M
Trugnan
G
G
D000116:Acetylgalactosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001593:Benzyl Compounds / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics / Q000378:metabolism*; D004285:Dogs; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000201:enzymology; D007668:Kidney; D051379:Mice; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D019459:Urothelium / Q000166:cytology / Q000201:enzymology
D001593:Benzyl Compounds; D011994:Recombinant Proteins; C080739:benzyl-alpha-N-acetylgalactosamine; D018819:Dipeptidyl Peptidase 4; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine