Environmental Science and Pollution Research

, Volume 23, Issue 3, pp 2119–2127 | Cite as

Mechanistic insights into the specificity of human cytosolic sulfotransferase 2A1 (hSULT2A1) for hydroxylated polychlorinated biphenyls through the use of fluoro-tagged probes

  • E. J. Ekuase
  • T. J. van ‘t Erve
  • A. Rahaman
  • L. W. Robertson
  • M. W. Duffel
  • G. Luthe
PCBs: Exposures, Effects, Remediation and Regulation with special reference to PCBs in Schools


Determining the relationships between the structures of substrates and inhibitors and their interactions with drug-metabolizing enzymes is of prime importance in predicting the toxic potential of new and legacy xenobiotics. Traditionally, quantitative structure activity relationship (QSAR) studies are performed with many distinct compounds. Based on the chemical properties of the tested compounds, complex relationships can be established so that models can be developed to predict toxicity of novel compounds. In this study, the use of fluorinated analogues as supplemental QSAR compounds was investigated. Substituting fluorine induces changes in electronic and steric properties of the substrate without substantially changing the chemical backbone of the substrate. In vitro assays were performed using purified human cytosolic sulfotransferase hSULT2A1 as a model enzyme. A mono-hydroxylated polychlorinated biphenyl (4-OH PCB 14) and its four possible mono-fluoro analogues were used as test compounds. Remarkable similarities were found between this approach and previously published QSAR studies for hSULT2A1. Both studies implicate the importance of dipole moment and dihedral angle as being important to PCB structure in respect to being substrates for hSULT2A1. We conclude that mono-fluorinated analogues of a target substrate can be a useful tool to study the structure activity relationships for enzyme specificity.


F-tagged probes QSAR hSULT2A1 Polychlorinated biphenyls Hydroxylated polychlorinated biphenyls Sulfotransferase Computational chemistry 4-Hydroxy-3,5-dichlorobiphenyl 



This work was supported by the National Institutes of Health through research grants R01 CA038683 and P42ES 013661. We also acknowledge programmatic support through the University of Iowa Environmental Health Sciences Research Center (NIH grant P30 ES05605). Partial support was provided to Dr. Gregor Luthe by the Alexander von Humboldt Foundation. The project was financially supported by the Tech for Future fund, an initiative of the Saxion and Windesheim Universities of Applied Sciences and the regional government Overijsel, The Netherlands. The contents of this paper are solely the responsibility of the authors and do not necessarily represent the official views of the granting agencies.

Conflict of Interest

The authors state that no conflict of interest exists.


  1. Bensadoun A, Weinstein D (1976) Assay of proteins in the presence of interfering materials. Anal Biochem 70:241–250CrossRefGoogle Scholar
  2. Bergman A, Klasson-Wehler E, Kuroki H (1994) Selective retention of hydroxylated PCB metabolites in blood. Environ Health Perspect 102:464–469CrossRefGoogle Scholar
  3. Brouwer A, Longnecker MP, Birnbaum LS, Cogliano J, Kostyniak P, Moore J, Schantz S, Winneke G (1999) Characterization of potential endocrine-related health effects at low-dose levels of exposure to PCBs. Environ Health Perspect 107(Suppl 4):639–649CrossRefGoogle Scholar
  4. Cancès E, Mennucci B, Tomasi J (1997) A new integral equation formalism for the polarizable continuum model: theoretical background and applications to isotropic and anisotropic dielectrics. J Chem Phys 107:3032CrossRefGoogle Scholar
  5. Cossi M, Barone V, Cammi R, Tomasi J (1996) Ab initio study of solvated molecules: a new implementation of the polarizable continuum model. Chem Phys Lett 255:327–335CrossRefGoogle Scholar
  6. Currado GM, Harrad S (1998) Comparison of polychlorinated biphenyl concentrations in indoor and outdoor air and the potential significance of inhalation as a human exposure pathway. Environ Sci Technol 32:3043–3047CrossRefGoogle Scholar
  7. Dewar MJS, Zoebisch EG, Healy EF, Stewart JJP (1985) The development and use of quantum-mechanical molecular-models. 76. Am1 - a new general-purpose quantum-mechanical molecular-model. J Am Chem Soc 107:3902–3909CrossRefGoogle Scholar
  8. Duffel MW (2010) Sulfotransferases. Comprehensive toxicology, 4. Elsevier, OxfordGoogle Scholar
  9. Ekuase EJ, Liu Y, Lehmler HJ, Robertson LW, Duffel MW (2011) Structure-activity relationships for hydroxylated polychlorinated biphenyls as inhibitors of the sulfation of dehydroepiandrosterone catalyzed by human hydroxysteroid sulfotransferase SULT2A1. Chem Res Toxicol 24:1720–1728CrossRefGoogle Scholar
  10. Ekuase EJ, Lehmler HJ, Robertson LW, Duffel MW (2014) Binding interactions of hydroxylated polychlorinated biphenyls (OHPCBs) with human hydroxysteroid sulfotransferase hSULT2A1. Chem Biol Interact 212:56–64CrossRefGoogle Scholar
  11. Erickson MD, Kaley RG 2nd (2011) Applications of polychlorinated biphenyls. Environ Sci Pollut Res Int 18:135–151CrossRefGoogle Scholar
  12. Gamage N, Barnett A, Hempel N, Duggleby RG, Windmill KF, Martin JL, McManus ME (2006) Human sulfotransferases and their role in chemical metabolism. Toxicol Sci 90:5–22CrossRefGoogle Scholar
  13. Grimm FA, Lehmler HJ, He X, Robertson LW, Duffel MW (2013) Sulfated metabolites of polychlorinated biphenyls are high-affinity ligands for the thyroid hormone transport protein transthyretin. Environ Health Perspect 121:657–662CrossRefGoogle Scholar
  14. Grimm FA, Hu D, Kania-Korwel I, Lehmler HJ, Ludewig G, Hornbuckle KC, Duffel MW, Bergman A, Robertson LW (2015) Metabolism and metabolites of polychlorinated biphenyls. Crit Rev Toxicol 45:245–272CrossRefGoogle Scholar
  15. Gulcan HO, Duffel MW (2011) Substrate inhibition in human hydroxysteroid sulfotransferase SULT2A1: studies on the formation of catalytically non-productive enzyme complexes. Arch Biochem Biophys 507:232–240CrossRefGoogle Scholar
  16. Gulcan HO, Liu Y, Duffel MW (2008) Pentachlorophenol and other chlorinated phenols are substrates for human hydroxysteroid sulfotransferase hSULT2A1. Chem Res Toxicol 21:1503–1508CrossRefGoogle Scholar
  17. Hansen LG (1998) Stepping backward to improve assessment of PCB congener toxicities. Environ Health Perspect 106(Suppl 1):171–189CrossRefGoogle Scholar
  18. Hawkins GD, Cramer CJ, Truhlar DG (1998) Universal quantum mechanical model for solvation free energies based on gas-phase geometries. J Phys Chem B 102:3257–3271CrossRefGoogle Scholar
  19. Herrick RF, McClean MD, Meeker JD, Baxter LK, Weymouth GA (2004) An unrecognized source of PCB contamination in schools and other buildings. Environ Health Perspect 112:1051–1053CrossRefGoogle Scholar
  20. Hu D, Hornbuckle KC (2010) Inadvertent polychlorinated biphenyls in commercial paint pigments. Environ Sci Technol 44:2822–2827CrossRefGoogle Scholar
  21. James MO (2001) Polychlorinated biphenyls: metabolism and metabolites. PCBs: recent advances in environmental toxicology and health effects. The University Press of Kentucky, LexingtonGoogle Scholar
  22. James MO, Ambadapadi S (2013) Interactions of cytosolic sulfotransferases with xenobiotics. Drug Metab Rev 45:401–414CrossRefGoogle Scholar
  23. Kester MH, Bulduk S, Tibboel D, Meinl W, Glatt H, Falany CN, Coughtrie MW, Bergman A, Safe SH, Kuiper GG, Schuur AG, Brouwer A, Visser TJ (2000) Potent inhibition of estrogen sulfotransferase by hydroxylated PCB metabolites: a novel pathway explaining the estrogenic activity of PCBs. Endocrinology 141:1897–1900CrossRefGoogle Scholar
  24. Kim JS, Klosener J, Flor S, Peters TM, Ludewig G, Thorne PS, Robertson LW, Luthe G (2014) Toxicity assessment of air-delivered particle-bound polybrominated diphenyl ethers. Toxicology 317:31–39CrossRefGoogle Scholar
  25. Klösener J, Swenson DC, Robertson LW, Luthe G (2008) Effects of fluoro substitution on 4-bromodiphenyl ether (PBDE 3). Acta Crystallogr B 64:108–119CrossRefGoogle Scholar
  26. Klösener J, Peters TM, Adamcakova-Dodd A, Teesch LM, Thorne PS, Robertson LW, Luthe G (2009) Innovative application of fluoro tagging to trace airborne particulate and gas-phase polybrominated diphenyl ether exposures. Chem Res Toxicol 22:179–186CrossRefGoogle Scholar
  27. Lauby-Secretan B, Loomis D, Grosse Y, El Ghissassi F, Bouvard V, Benbrahim-Tallaa L, Guha N, Baan R, Mattock H, Straif K, International Agency for Research on Cancer Monograph Working Group Iarc LF (2013) Carcinogenicity of polychlorinated biphenyls and polybrominated biphenyls. Lancet Oncol 14:287–288CrossRefGoogle Scholar
  28. Liu Y, Apak TI, Lehmler HJ, Robertson LW, Duffel MW (2006) Hydroxylated polychlorinated biphenyls are substrates and inhibitors of human hydroxysteroid sulfotransferase SULT2A1. Chem Res Toxicol 19:1420–1425CrossRefGoogle Scholar
  29. Liu Y, Smart JT, Song Y, Lehmler HJ, Robertson LW, Duffel MW (2009) Structure-activity relationships for hydroxylated polychlorinated biphenyls as substrates and inhibitors of rat sulfotransferases and modification of these relationships by changes in thiol status. Drug Metab Dispos 37:1065–1072CrossRefGoogle Scholar
  30. Ludewig G, Robertson LW (2013) Polychlorinated biphenyls (PCBs) as initiating agents in hepatocellular carcinoma. Cancer Lett 334:46–55CrossRefGoogle Scholar
  31. Luthe G, Brinkman UA (2000) Monofluorinated polycyclic aromatic hydrocarbons: characteristics and intended use in environmental analysis. Analyst 125:1699–1702CrossRefGoogle Scholar
  32. Luthe G, Ariese F, Brinkman UAT (2002a) Monofluorinated polycyclic aromatic hydrocarbons: standards in environmental chemistry and biochemical applications. In: Neilson AH (ed) Handbook of environmental chemistry: organic fluorine compounds. Springer Verlag, BerlinGoogle Scholar
  33. Luthe G, Stroomberg GJ, Ariese F, Brinkman UA, van Straalen NM (2002b) Metabolism of 1-fluoropyrene and pyrene in marine flatfish and terrestrial isopods. Environ Toxicol Pharmacol 12:221–229CrossRefGoogle Scholar
  34. Luthe G, Leonards PE, Reijerink GS, Liu H, Johansen JE, Robertson LW (2006) Monofluorinated analogues of polybrominated diphenyl ethers as analytical standards: synthesis, NMR, and GC-MS characterization and molecular orbital studies. Environ Sci Technol 40:3023–3029CrossRefGoogle Scholar
  35. Luthe G, Swenson DC, Robertson LW (2007) Influence of fluoro-substitution on the planarity of 4-chlorobiphenyl (PCB 3). Acta Crystallogr B 63:319–327CrossRefGoogle Scholar
  36. Luthe G, Garcia Boy R, Jacobus J, Smith BJ, Rahaman A, Robertson LW, Ludewig G (2008a) Xenobiotic geometry and media pH determine cytotoxicity through solubility. Chem Res Toxicol 21:1017–1027CrossRefGoogle Scholar
  37. Luthe G, Jacobus JA, Robertson LW (2008b) Receptor interactions by polybrominated diphenyl ethers versus polychlorinated biphenyls: a theoretical structure-activity assessment. Environ Toxicol Pharmacol 25:202–210CrossRefGoogle Scholar
  38. Luthe GM, Schut BG, Aaseng JE (2009) Monofluorinated analogues of polychlorinated biphenyls (F-PCBs): synthesis using the Suzuki-coupling, characterization, specific properties and intended use. Chemosphere 77:1242–1248CrossRefGoogle Scholar
  39. Marek RF, Martinez A, Hornbuckle KC (2013) Discovery of hydroxylated polychlorinated biphenyls (OH-PCBs) in sediment from a lake Michigan waterway and original commercial aroclors. Environ Sci Technol 47:8204–8210CrossRefGoogle Scholar
  40. Matthews HB, Kato S (1979) The metabolism and disposition of halogenated aromatics. Ann N Y Acad Sci 320:131–137CrossRefGoogle Scholar
  41. Pacifici GM, Coughtrie MW (2005) Human cytosolic sulfotransferases. CRC Press, Boca RatonGoogle Scholar
  42. Persoon C, Peters TM, Kumar N, Hornbuckle KC (2010) Spatial distribution of airborne polychlorinated biphenyls in Cleveland, Ohio and Chicago, Illinois. Environ Sci Technol 44:2797–2802CrossRefGoogle Scholar
  43. Pery AR, Desmots S, Mombelli E (2010) Substance-tailored testing strategies in toxicology: an in silico methodology based on QSAR modeling of toxicological thresholds and Monte Carlo simulations of toxicological testing. Regul Toxicol Pharmacol 56:82–92CrossRefGoogle Scholar
  44. Quinete N, Schettgen T, Bertram J, Kraus T (2014) Occurrence and distribution of PCB metabolites in blood and their potential health effects in humans: a review. Environ Sci Pollut Res Int 21:11951–11972CrossRefGoogle Scholar
  45. Sandau CD, Ayotte P, Dewailly E, Duffe J, Norstrom RJ (2000) Analysis of hydroxylated metabolites of PCBs (OH-PCBs) and other chlorinated phenolic compounds in whole blood from Canadian inuit. Environ Health Perspect 108:611–616CrossRefGoogle Scholar
  46. Schantz SL (1996) Developmental neurotoxicity of PCBs in humans: what do we know and where do we go from here? Neurotoxicol Teratol 18:217–227, discussion 229-76CrossRefGoogle Scholar
  47. Seegal RF (1996) Epidemiological and laboratory evidence of PCB-induced neurotoxicity. Crit Rev Toxicol 26:709–737CrossRefGoogle Scholar
  48. Sekura RD (1981) Adenosine 3′-phosphate 5′-phosphosulfate. Methods Enzymol 77:413–415CrossRefGoogle Scholar
  49. Sekura RD, Jakoby WB (1979) Phenol sulfotransferases. J Biol Chem 254:5658–5663Google Scholar
  50. Shao Y et al (2006) Advances in methods and algorithms in a modern quantum chemistry program package. Phys Chem Chem Phys 8:3172–3191CrossRefGoogle Scholar
  51. Sheng JJ, Duffel MW (2003) Enantioselectivity of human hydroxysteroid sulfotransferase ST2A3 with naphthyl-1-ethanols. Drug Metab Dispos 31:697–700CrossRefGoogle Scholar
  52. Sheng JJ, Sharma V, Duffel MW (2001) Measurement of aryl and alcohol sulfotransferase activity. Curr Protoc Toxicol Chapter 4, Unit4 5Google Scholar
  53. Soffers AE, Boersma MG, Vaes WH, Vervoort J, Tyrakowska B, Hermens JL, Rietjens IM (2001) Computer-modeling-based QSARs for analyzing experimental data on biotransformation and toxicity. Toxicol In Vitro 15:539–551CrossRefGoogle Scholar
  54. Ueno D, Darling C, Alaee M, Campbell L, Pacepavicius G, Teixeira C, Muir D (2007) Detection of hydroxylated polychlorinated biphenyls (OH-PCBs) in the abiotic environment: surface water and precipitation from Ontario, Canada. Environ Sci Technol 41:1841–1848CrossRefGoogle Scholar
  55. van ‘t Erve TJ, Rautiainen RH, Robertson LW, Luthe G (2010) Trimethylsilyldiazomethane: a safe non-explosive, cost effective and less-toxic reagent for phenol derivatization in GC applications. Environ Int 36:835–842CrossRefGoogle Scholar
  56. Winkler DA, Mombelli E, Pietroiusti A, Tran L, Worth A, Fadeel B, McCall MJ (2013) Applying quantitative structure-activity relationship approaches to nanotoxicology: current status and future potential. Toxicology 313:15–23CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • E. J. Ekuase
    • 1
  • T. J. van ‘t Erve
    • 2
    • 3
    • 4
    • 7
  • A. Rahaman
    • 5
  • L. W. Robertson
    • 2
    • 4
  • M. W. Duffel
    • 1
    • 4
  • G. Luthe
    • 2
    • 3
    • 4
    • 6
  1. 1.Department of Pharmaceutical Sciences and Experimental TherapeuticsThe University of IowaIowa CityUSA
  2. 2.Department of Occupational and Environmental HealthThe University of IowaIowa CityUSA
  3. 3.Institute of Life SciencesSaxion University of Applied SciencesEnschedeThe Netherlands
  4. 4.Interdisciplinary Graduate Program in Human ToxicologyThe University of IowaIowa CityUSA
  5. 5.Department of ChemistryThe University of IowaIowa CityUSA
  6. 6.LuthePharmaGronauGermany
  7. 7.Immunity, Inflammation and Disease LaboratoryNational Institute of Environmental Health SciencesResearch Triangle ParkUSA

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