N-glycosylation modifies prostaglandin E uptake by reducing cell surface expression of SLCO2A1.

SLCO2A1 functions as a prostaglandin (PG) influx transporter to facilitate intracellular oxidation of PGs and its defect causes dysregulation of PG signaling and metabolism. This study aimed to clarify effects of N-glycosylation on functional SLCO2A1 expression. Putative N-glycosylation site(s) (N134, N478, and/or N491) of human SLCO2A1 were mutated to Q and wild-type (WT) and mutant forms were expressed in HEK293 and human epithelial cells. Molecular weight of WT decreased to nearly 55 kDa by PNGase F treatment and was identical to that of triple mutant (TM, i.e., N134Q/N478Q/N491Q). Transport affinity of TM for PGE (K of 392.7 nM) was comparable to that of WT (K of 328.5 nM); however, immunoassays showed that TM cell surface expression remained at 24% of WT in HEK293 cells, resulting in a reduced cellular PGE uptake. These results suggest N-glycosylation modifies cellular PGE uptake by decreasing SLCO2A1 localization to the plasma membrane.