Archives of Toxicology

, Volume 90, Issue 1, pp 217–227 | Cite as

Interaction of perfluoroalkyl acids with human liver fatty acid-binding protein

  • Nan Sheng
  • Juan Li
  • Hui Liu
  • Aiqian ZhangEmail author
  • Jiayin DaiEmail author
Organ Toxicity and Mechanisms


Perfluoroalkyl acids (PFAAs) are highly persistent and bioaccumulative, resulting in their broad distribution in humans and the environment. The liver is an important target for PFAAs, but the mechanisms behind PFAAs interaction with hepatocyte proteins remain poorly understood. We characterized the binding of PFAAs to human liver fatty acid-binding protein (hL-FABP) and identified critical structural features in their interaction. The binding interaction of PFAAs with hL-FABP was determined by fluorescence displacement and isothermal titration calorimetry (ITC) assay. Molecular simulation was conducted to define interactions at the binding sites. ITC measurement revealed that PFOA/PFNA displayed a moderate affinity for hL-FABP at a 1:1 molar ratio, a weak binding affinity for PFHxS and no binding for PFHxA. Moreover, the interaction was mainly mediated by electrostatic attraction and hydrogen bonding. Substitution of Asn111 with Asp caused loss of binding affinity to PFAA, indicating its crucial role for the initial PFAA binding to the outer binding site. Substitution of Arg122 with Gly caused only one molecule of PFAA to bind to hL-FABP. Molecular simulation showed that substitution of Arg122 increased the volume of the outer binding pocket, making it impossible to form intensive hydrophobic stacking and hydrogen bonds with PFOA, and highlighting its crucial role in the binding process. The binding affinity of PFAAs increased significantly with their carbon number. Arg122 and Asn111 played a pivotal role in these interactions. Our findings may help understand the distribution pattern, bioaccumulation, elimination, and toxicity of PFAAs in humans.


Perfluorinated compounds Human liver fatty acid-binding protein Interaction Isothermal titration calorimetry Molecular simulation 



This work was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB14000000) and the National Natural Science Foundation of China (grants 31320103915 and 31025006 for Dai, 21277164 for Zhang).

Conflict of interest

The authors declare they have no competing financial interests.

Supplementary material

204_2014_1391_MOESM1_ESM.docx (2.3 mb)
Supplementary material 1 (DOCX 2340 kb)


  1. Atshaves BP, Martin GG, Hostetler HA, McIntosh AL, Kier AB, Schroeder F (2010) Liver fatty acid-binding protein and obesity. J Nutr Biochem 21:1015–1032PubMedPubMedCentralCrossRefGoogle Scholar
  2. Bernlohr DA, Simpson MA, Hertzel AV, Banaszak LJ (1997) Intracellular lipid-binding proteins and their genes. Annu Rev Nutr 17:277–303PubMedCrossRefGoogle Scholar
  3. Carbone V, Velkov T (2013) Interaction of phthalates and phenoxy acid herbicide environmental pollutants with intestinal intracellular lipid binding proteins. Chem Res Toxicol 26:1240–1250PubMedCrossRefGoogle Scholar
  4. De Silva AO, Mabury SA (2006) Isomer distribution of perfluorocarboxylates in human blood: potential correlation to source. Environ Sci Technol 40:2903–2909PubMedCrossRefGoogle Scholar
  5. Ding L, Hao F, Shi Z, Wang Y, Zhang H, Tang H et al (2009) Systems biological responses to chronic perfluorododecanoic acid exposure by integrated metabonomic and transcriptomic studies. J Proteome Res 8:2882–2891PubMedCrossRefGoogle Scholar
  6. Ehresman DJ, Froehlich JW, Olsen GW, Chang SC, Butenhoff JL (2007) Comparison of human whole blood, plasma, and serum matrices for the determination of perfluorooctanesulfonate (pfos), perfluorooctanoate (pfoa), and other fluorochemicals. Environ Res 103:176–184PubMedCrossRefGoogle Scholar
  7. Fang X, Zou S, Zhao Y, Cui R, Zhang W, Hu J et al (2012) Kupffer cells suppress perfluorononanoic acid-induced hepatic peroxisome proliferator-activated receptor alpha expression by releasing cytokines. Arch Toxicol 86:1515–1525PubMedCrossRefGoogle Scholar
  8. Hostetler HA, McIntosh AL, Atshaves BP, Storey SM, Payne HR, Kier AB et al (2009) L-fabp directly interacts with pparalpha in cultured primary hepatocytes. J Lipid Res 50:1663–1675PubMedPubMedCentralCrossRefGoogle Scholar
  9. Kannan K, Corsolini S, Falandysz J, Fillmann G, Kumar KS, Loganathan BG et al (2004) Perfluorooctanesulfonate and related fluorochemicals in human blood from several countries. Environ Sci Technol 38:4489–4495PubMedCrossRefGoogle Scholar
  10. Kelly SM, Jess TJ, Price NC (2005) How to study proteins by circular dichroism. Biochim Biophys Acta 1751:119–139PubMedCrossRefGoogle Scholar
  11. Kennedy GL Jr, Butenhoff JL, Olsen GW, O’Connor JC, Seacat AM, Perkins RG et al (2004) The toxicology of perfluorooctanoate. Crit Rev Toxicol 34:351–384PubMedCrossRefGoogle Scholar
  12. Krafft MP, Riess JG (2009) Chemistry, physical chemistry, and uses of molecular fluorocarbon–hydrocarbon diblocks, triblocks, and related compounds–unique “apolar” components for self-assembled colloid and interface engineering. Chem Rev 109:1714–1792PubMedCrossRefGoogle Scholar
  13. Lau C, Anitole K, Hodes C, Lai D, Pfahles-Hutchens A, Seed J (2007) Perfluoroalkyl acids: a review of monitoring and toxicological findings. Toxicol Sci 99:366–394PubMedCrossRefGoogle Scholar
  14. Lawrence JW, Kroll DJ, Eacho PI (2000) Ligand-dependent interaction of hepatic fatty acid-binding protein with the nucleus. J Lipid Res 41:1390–1401PubMedGoogle Scholar
  15. Luebker DJ, Hansen KJ, Bass NM, Butenhoff JL, Seacat AM (2002) Interactions of fluorochemicals with rat liver fatty acid-binding protein. Toxicology 176:175–185PubMedCrossRefGoogle Scholar
  16. Maestri L, Negri S, Ferrari M, Ghittori S, Fabris F, Danesino P et al (2006) Determination of perfluorooctanoic acid and perfluorooctanesulfonate in human tissues by liquid chromatography/single quadrupole mass spectrometry. Rapid Commun Mass Spectrom 20:2728–2734PubMedCrossRefGoogle Scholar
  17. Olsen GW, Burris JM, Ehresman DJ, Froehlich JW, Seacat AM, Butenhoff JL et al (2007) Half-life of serum elimination of perfluorooctanesulfonate, perfluorohexanesulfonate, and perfluorooctanoate in retired fluorochemical production workers. Environ Health Perspect 115:1298–1305PubMedPubMedCentralCrossRefGoogle Scholar
  18. Renner R (2001) Growing concern over perfluorinated chemicals. Environ Sci Technol 35:154A–160APubMedCrossRefGoogle Scholar
  19. Sakr CJ, Leonard RC, Kreckmann KH, Slade MD, Cullen MR (2007) Longitudinal study of serum lipids and liver enzymes in workers with occupational exposure to ammonium perfluorooctanoate. J Occup Environ Med 49:872–879PubMedCrossRefGoogle Scholar
  20. Sharma A, Sharma A (2011) Fatty acid induced remodeling within the human liver fatty acid-binding protein. J Biol Chem 286:31924–31928PubMedPubMedCentralCrossRefGoogle Scholar
  21. Sreerama N, Woody RW (2000) Estimation of protein secondary structure from circular dichroism spectra: comparison of contin, selcon, and cdsstr methods with an expanded reference set. Anal Biochem 287:252–260PubMedCrossRefGoogle Scholar
  22. Steenland K, Tinker S, Frisbee S, Ducatman A, Vaccarino V (2009) Association of perfluorooctanoic acid and perfluorooctane sulfonate with serum lipids among adults living near a chemical plant. Am J Epidemiol 170:1268–1278PubMedCrossRefGoogle Scholar
  23. Vanden Heuvel JP, Thompson JT, Frame SR, Gillies PJ (2006) Differential activation of nuclear receptors by perfluorinated fatty acid analogs and natural fatty acids: a comparison of human, mouse, and rat peroxisome proliferator-activated receptor-alpha, -beta, and -gamma, liver × receptor-beta, and retinoid × receptor-alpha. Toxicol Sci 92:476–489PubMedCrossRefGoogle Scholar
  24. Velkov T, Chuang S, Wielens J, Sakellaris H, Charman WN, Porter CJ et al (2005) The interaction of lipophilic drugs with intestinal fatty acid-binding protein. J Biol Chem 280:17769–17776PubMedCrossRefGoogle Scholar
  25. Weiss J, Andersson P, Lamoree M, Leonards P, van Leeuwen S, Hamers T (2009) Competitive binding of poly- and perfluorinated compounds to the thyroid hormone transport protein transthyretin. Toxicol Sci 109:206–216PubMedCrossRefGoogle Scholar
  26. Wolfrum C, Borrmann CM, Borchers T, Spener F (2001) Fatty acids and hypolipidemic drugs regulate peroxisome proliferator-activated receptors alpha- and gamma-mediated gene expression via liver fatty acid binding protein: a signaling path to the nucleus. Proc Natl Acad Sci U S A 98:2323–2328PubMedPubMedCentralCrossRefGoogle Scholar
  27. Zhang W, Zhang Y, Taniyasu S, Yeung LW, Lam PK, Wang J et al (2013) Distribution and fate of perfluoroalkyl substances in municipal wastewater treatment plants in economically developed areas of china. Environ Pollut 176:10–17PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Key Laboratory of Animal Ecology and Conservation Biology, Institute of ZoologyChinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental SciencesChinese Academy of SciencesBeijingPeople’s Republic of China

Personalised recommendations