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Cited by 3Drug Transport and Resistance MechanismsReceptor Mechanisms and SignalingIon channel regulation and function

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Structure and post-translational modification of the prostaglandin transporter

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Yu, Peixuan, Orlando, Melanie A., Orlando, Benjamin J.·2025·Communications Biology
DOI 10.1038/s42003-025-08510-0PMC PMC12274398

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Abstract

Abstract The prostaglandin transporter (PGT) is a member of the Organic Anion Transporting Polypeptide (OATP) family of membrane transporters. PGT mediates the uptake of prostaglandins from the extracellular environment to enable intracellular enzymatic degradation and termination of signaling. In addition to importing prostaglandins, PGT is also an essential core component of the Maxi-Cl channel, which facilitates cellular release of ATP and other small organic anions. Despite progress on understanding the (patho)physiological roles of PGT, and development of small molecules to inhibit this transporter, molecular details of the overall structure and transport mechanism remain elusive. Here we determined the cryo-EM structure of human PGT, which demonstrates an overall topology consistent with other OATPs despite possessing a dual transporter/channel functionality. We additionally investigated the role of eight potential disulfide bonds found in the extracellular loops of PGT and paralogous transporters. Through biochemical and functional characterization we demonstrate that six intra-loop disulfide bonds (C420-C511, C450-C470, C492-C474, C459-C507, C444-C494, C580-C594) are essential for proper N-glycosylation, plasma membrane trafficking, and prostaglandin import activity of PGT. In contrast, two inter-loop disulfides (C155-C587 and C143-C448) were found to restrict maximal prostaglandin uptake, suggesting a possible regulatory role in modulating PGT activity. In total, our studies provide a fresh molecular perspective on the structure, post-translational modification, and overall function of PGT. Significance Statement The prostaglandin transporter (PGT) is essential for cellular uptake of prostaglandins and serves as a core component of the Maxi-Cl channel. Using cryo-EM we resolved the structure of human PGT, revealing a similar overall topology as other OATP transporters. Extensive site-directed mutagenesis and functional assays identified eight disulfide bonds in PGT’s extracellular domain as key regulators of glycosylation, trafficking, and transport activity. These findings provide a structural basis for PGT function and lay a foundation to further explore substrate recognition and inhibitor design for this biologically significant transporter.

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Research works printed but not linked: 58

  1. Ricciotti, E. & Fitzgerald, G. A. Prostaglandins and inflammation. Arterioscler. Thromb. Vasc. Biol. 31, 986–1000 (2011). · DOI
  2. Yerushalmi, G. M. et al. The prostaglandin transporter (PGT) as a potential mediator of ovulation. Sci. Transl. Med. 8, 338ra68 (2016). · DOI
  3. Niringiyumukiza, J. D., Cai, H. & Xiang, W. Prostaglandin E2 involvement in mammalian female fertility: ovulation, fertilization, embryo development and early implantation. Reprod. Biol. Endocrinol. 16, 43 (2018). · DOI
  4. Chi, Y. et al. Inhibition of the prostaglandin transporter PGT lowers blood pressure in hypertensive rats and mice. PLoS One 10, e0131735 (2015). · DOI
  5. Finetti, F. et al. Prostaglandin E2 and cancer: insight into tumor progression and immunity. Biology9, 1–26 (2020). · DOI
  6. Reid, G. et al. The human multidrug resistance protein MRP4 functions as a prostaglandin efflux transporter and is inhibited by nonsteroidal antiinflammatory drugs. Proc. Natl Acad. Sci. USA 100, 9244–9249 (2003). · DOI · Paper page
  7. Pourmal, S. et al. Structural basis of prostaglandin efflux by MRP4. Nat. Struct. Mol. Biol. 31, 621–632 (2024). · DOI · Paper page
  8. Bloch, M., Raj, I., Pape, T. & Taylor, N. M. I. Structural and mechanistic basis of substrate transport by the multidrug transporter MRP4. Structure 31, 1407–1418.e6 (2023). · DOI · Paper page
  9. Sugimoto, Y. & Narumiya, S. Prostaglandin E receptors. J. Biol. Chem. 282, 11613–11617 (2007). · DOI
  10. Nomura, T., Lu, R., Pucci, M. L. & Schuster, V. L. The two-step model of prostaglandin signal termination: in vitro reconstitution with the prostaglandin transporter and prostaglandin 15 dehydrogenase. Mol. Pharm. 65, 973–978 (2004). · DOI
  11. Nakanishi, T., Nakamura, Y. & Umeno, J. Recent advances in studies of SLCO2A1 as a key regulator of the delivery of prostaglandins to their sites of action. Pharm. Ther. 223, 107803 (2021). · DOI
  12. Kanai, N. et al. Identification and characterization of a prostaglandin transporter. N. Ser. 268, 866–869 (1995).
  13. Nakata, R. et al. Slco2a1 deficiency exacerbates experimental colitis via inflammasome activation in macrophages: a possible mechanism of chronic enteropathy associated with SLCO2A1 gene. Sci. Rep. 10, 4883 (2020). · DOI
  14. Inagaki, M. et al. Contribution of prostaglandin transporter OATP2A1/SLCO2A1 to placenta-to-maternal hormone signaling and labor induction. iScience 23, 101098 (2020). · DOI
  15. Chang, H. Y., Locker, J., Lu, R. & Schuster, V. L. Failure of postnatal ductus arteriosus closure in prostaglandin transporter-deficient mice. Circulation 121, 529–536 (2010). · DOI
  16. Zhang, Z. et al. Exome sequencing identifies SLCO2A1 mutations as a cause of primary hypertrophic osteoarthropathy. Am. J. Hum. Genet 90, 125–132 (2012). · DOI
  17. Umeno, J. et al. A hereditary enteropathy caused by mutations in the SLCO2A1 gene, encoding a prostaglandin transporter. PLoS Genet 11, e1005581 (2015). · DOI
  18. Liu, Z., Benard, O., Syeda, M. M., Schuster, V. L. & Chi, Y. Inhibition of prostaglandin transporter (PGT) promotes perfusion and vascularization and accelerates wound healing in non-diabetic and diabetic rats. PLoS One 10, e0133615 (2015). · DOI
  19. Chi, Y. et al. Development of a high-affinity inhibitor of the prostaglandin transporter. J. Pharmacol. Exp. Therapeutics 339, 633–641 (2011). · DOI
  20. Islam, M. R. et al. Annexin A2-S100A10 represents the regulatory component of Maxi-Cl channel dependent on protein tyrosine dephosphorylation and intracellular Ca2. Cell. Physiol. Biochem. 54, 538–555 (2020). · DOI
  21. Sabirov, R. Z. et al. The organic anion transporter SLCO 2A1 constitutes the core component of the Maxi-Cl channel. EMBO J. 36, 3309–3324 (2017). · DOI
  22. Murray, M. & Zhou, F. Trafficking and other regulatory mechanisms for organic anion transporting polypeptides and organic anion transporters that modulate cellular drug and xenobiotic influx and that are dysregulated in disease. Br. J. Pharm. 174, 1908–1924 (2017). · DOI
  23. Hänggi, E., Grundschober, A. F., Leuthold, S., Meier, P. J. & St Pierre, M. V. Functional analysis of the extracellular cysteine residues in the human organic anion transporting polypeptide, OATP2B1. Mol. Pharm. 70, 806–817 (2006). · DOI
  24. Shan, Z. et al. Cryo-EM structures of human organic anion transporting polypeptide OATP1B1. Cell Res. 33, 940–951 (2023). · DOI
  25. Ciută, A. D. et al. Structure of human drug transporters OATP1B1 and OATP1B3. Nat. Commun. 14, 5774 (2023). · DOI
  26. Diggle, C. P. et al. Prostaglandin transporter mutations cause pachydermoperiostosis with myelofibrosis. Hum. Mutat. 33, 1175–1181 (2012). · DOI
  27. Jiang, Y. et al. Novel SLCO2A1compound heterozygous mutation causing primary hypertrophic osteoarthropathy with Bartter-like hypokalemia in a Chinese family. J. Endocrinol. Invest. 42, 1245–1252 (2019). · DOI
  28. Sonoda, A. et al. Characteristic facial appearance was the key to diagnosing chronic enteropathy associated with SLCO2A1-associated primary hypertrophic osteoarthropathy. Intern. Med. 59, 491–494 (2020). · DOI
  29. Yuan, L. et al. Novel slco2a1 mutations cause gender-differentiated pachydermoperiostosis. Endocr. Connect 7, 1116–1128 (2018). · DOI
  30. Tanaka, K., Zhou, F., Kuze, K. & You, G. Cysteine residues in the organic anion transporter mOAT1. Biochem. J. 380, 283–287 (2004). · DOI
  31. Nakamura, Y., Aizawa, C., Kawata, H. & Nakanishi, T. N-glycosylation modifies prostaglandin E2 uptake by reducing cell surface expression of SLCO2A1. Prostaglandins Other Lipid Mediat 165, 106714 (2023). · DOI
  32. Eberhardt, J., Santos-Martins, D., Tillack, A. F. & Forli, S. AutoDock Vina 1.2.0: new docking methods, expanded force field, and python bindings. J. Chem. Inf. Model 61, 3891–3898 (2021). · DOI
  33. Chan, B. S., Satriano, J. A. & Schuster, V. L. Mapping the substrate binding site of the prostaglandin transporter PGT by cysteine scanning mutagenesis. J. Biol. Chem. 274, 25564–25570 (1999). · DOI
  34. Chan, B. S., Bao, Y. & Schuster, V. L. Role of conserved transmembrane cationic amino acids in the prostaglandin transporter PGT. Biochemistry 41, 9215–9221 (2002). · DOI
  35. Schlott, B. et al. Interaction of Kazal-type inhibitor domains with serine proteinases: biochemical and structural studies. J. Mol. Biol. 318, 533–546 (2002). · DOI
  36. Li, Y. et al. Importance of N-glycosylation for the expression and function of human organic anion transporting polypeptide 2B1. ACS Pharm. Transl. Sci. 6, 1347–1356 (2023). · DOI
  37. Nakamura, Y. et al. Prostaglandin transporter OATP2A1/SLCO2A1 is essential for body temperature regulation during fever. J. Neurosci. 38, 5584–5595 (2018). · DOI
  38. Kamo, S. et al. Impact of FDA-approved drugs on the prostaglandin transporter OATP2A1/SLCO2A1. J. Pharm. Sci. 106, 2483–2490 (2017). · DOI
  39. Nakamura, Y., Kozakai, H., Nishio, T., Yoshida, K. & Nakanishi, T. Phenolsulfonphthalein as a surrogate substrate to assess altered function of the prostaglandin transporter SLCO2A1. Drug Metab Pharmacokinet 44, 100452 (2022). · DOI
  40. Matsuo, Y. & Yodoi, J. Extracellular thioredoxin: a therapeutic tool to combat inflammation. Cytokine Growth Factor Rev. 24, 345–353 (2013). · DOI
  41. Lei, Y. et al. Redox regulation of inflammation: old elements, a new story. Med. Res. Rev. 35, 306–340 (2015). · DOI
  42. Yi, M. C. & Khosla, C. Thiol-disulfide exchange reactions in the mammalian extracellular environment. Annu. Rev. Chem. Biomol. Eng. 7, 197–222 (2016). · DOI
  43. Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. CryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017). · DOI · Paper page
  44. Punjani, A., Zhang, H. & Fleet, D. J. Non-uniform refinement: adaptive regularization improves single-particle cryo-EM reconstruction. Nat. Methods 17, 1214–1221 (2020). · DOI
  45. Kimanius, D. et al. Data-driven regularization lowers the size barrier of cryo-EM structure determination. Nat. Methods 21, 1216–1221 (2024). · DOI
  46. Pettersen, E. F. et al. UCSF Chimera - a visualization system for exploratory research and analysis. J. Comput Chem. 25, 1605–1612 (2004). · DOI
  47. Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D. Biol. Crystallogr. 66, 486–501 (2010). · DOI · Paper page
  48. Adams, P. D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D. Biol. Crystallogr. 66, 213–221 (2010). · DOI · Paper page
  49. Williams, C. J. et al. MolProbity: more and better reference data for improved all-atom structure validation. Protein Sci. 27, 293–315 (2018). · DOI · Paper page
  50. Pettersen, E. F. et al. UCSF ChimeraX: structure visualization for researchers, educators, and developers. Protein Sci. 30, 70–82 (2021). · DOI
  51. Morris, G. M. et al. Software news and updates AutoDock4 and AutoDockTools4: automated docking with selective receptor flexibility. J. Comput. Chem. 30, 2785–2791 (2009). · DOI
  52. Feng, S., Park, S., Choi, Y. K. & Im, W. CHARMM-GUI membrane builder: past, current, and future developments and applications. J. Chem. Theory Comput 19, 2161–2185 (2023). · DOI
  53. Huang, J. et al. CHARMM36m: An improved force field for folded and intrinsically disordered proteins. Nat. Methods 14, 71–73 (2016). · DOI
  54. Izaguirre, J. A., Sweet, C. R. & Pande, V. S. Multiscale dynamics of macromolecules using normal mode Langevin. Biocomputing 2010, 240–251 (2009).
  55. Åqvist, J., Wennerström, P., Nervall, M., Bjelic, S. & Brandsdal, B. O. Molecular dynamics simulations of water and biomolecules with a Monte Carlo constant pressure algorithm. Chem. Phys. Lett. 384, 288–294 (2004). · DOI
  56. Essmann, U. et al. A smooth particle mesh Ewald method. J. Chem. Phys. 103, 8577–8593 (1995). · DOI
  57. Humphrey, W., Dalke, A. & Schulten, K. VMD: visual molecular dynamics. J. Mol. Graph 14, 33–38 (1996). · DOI · Paper page
  58. Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012). · DOI · Paper page

Metadata

Authors
Yu, P., Orlando, M. A., & Orlando, B. J.
Affiliations
Michigan State University (US)
Corresponding author
Orlando, Benjamin J.
ORCID
Orlando, Benjamin J.
Venue
Communications Biology
Publisher
Springer Science and Business Media LLC
Published
2025-07-18
Volume
8
Issue
1
Pages
1066-1066
ISSN
2399-3642 (ISSN-L)
DOI
10.1038/s42003-025-08510-0
PMC
PMC12274398
OpenAlex
W4412523165
Cited by
3
References
2
Citations by year
2026: 2 · 2025: 1
FWCI
1.24
Type
journal article
Language
en
Open access
Licence
Locations
  • Communications Biology (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Research reported in this publication was supported by departmental startup funds and the James K. Billman Jr. MD Endowed Research Professorship to B.J.O
Topics
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
Keywords
prostaglandin transporter, ATP release, disulfide bonds, N-glycosylation, plasma membrane trafficking, transporter structure, cryo-EM structure, transporter inhibition
MeSH terms
Disulfides, Glycosylation, Humans, Prostaglandins, Protein Conformation, Protein Processing, Post-Translational, Cryoelectron Microscopy, Organic Anion Transporters

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