N-glycosylation modifies prostaglandin E2 uptake by reducing cell surface expression of SLCO2A1

Prostaglandins Other Lipid Mediat. 2023 Apr:165:106714. doi: 10.1016/j.prostaglandins.2023.106714. Epub 2023 Jan 24.

Abstract

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 PGE2 (Km of 392.7 nM) was comparable to that of WT (Km 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 PGE2 uptake. These results suggest N-glycosylation modifies cellular PGE2 uptake by decreasing SLCO2A1 localization to the plasma membrane.

Keywords: N-glycosylation; PGE(2); Prostaglandin transporter; SLCO2A1; Subcellular localization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport
  • Cell Membrane / metabolism
  • Dinoprostone* / metabolism
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Organic Anion Transporters* / genetics
  • Organic Anion Transporters* / metabolism

Substances

  • Dinoprostone
  • SLCO2A1 protein, human
  • Organic Anion Transporters