Interaction of curcumin with human serum albumin—A spectroscopic study

Abstract

Curcumin (diferuloyl methane) has a wide range of physiological and pharmacological actions. Curcumin interaction with human serum albumin (HSA) has been followed by fluorescence quenching and circular dichroism (CD) measurements. Based on fluorescence measurements, the equilibrium constant for the interaction is 2.0±0.2×105M−1. Binding of curcumin to HSA induces an extrinsic CD band in the visible region. From the induced CD band measurements, the equilibrium constant has a value of 2.1±0.3×104M−1. Thus, HSA has two kinds of affinity sites for curcumin, one with high affinity and the other with lower affinity. Job’s plot indicated a binding stoichiometry of 1∶1 for the high-affinity site. The equilibrium constant was invariant with temperature in the range of 15 to 45°C, suggesting the role of hydrophobic interactions in the binding of curcumin to HSA. Curcumin does not change the conformation of the HSA molecule. These measurements have implications in the understanding of the curcumin transport under physiological conditions.

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Abbreviations

CD:

circular dichroism

HSA:

human serum albumin

Q max :

maximal quench

UV:

ultraviolet

References

  1. 1.

    Govindarajan, V.S. (1980) Turmeric-Chemistry, Technology and Quality, Crit. Rev. Food Sci. Nutr. 12, 199–301.

    PubMed  CAS  Article  Google Scholar 

  2. 2.

    Srimal, R.C., and Chawan, B.N. (1973) Pharmacology of Diferuloyl Methane (curcumin), a Nonsteroidal Anti-Inflammatory Agent. J. Pharm. Pharmacol. 25, 447–457.

    PubMed  CAS  Google Scholar 

  3. 3.

    Toda, S., Miyase, T., Arichi, H., Tanizawa, H., and Takino, Y. (1985) Natural Antioxidants. III. Antioxidative Components Isolated from Rhizome of Curcuma longa, L., Chem. Pharm. Bull. 33, 1725–1728.

    PubMed  CAS  Google Scholar 

  4. 4.

    Mukhopadhya, A., Basu, N., Ghatak, N., and Gujral, P.K. (1982) Anti-Inflammatory and Irritant Activities of Curcumin Analogues in Rats, Agents Actions 12, 508–515.

    Article  Google Scholar 

  5. 5.

    Pulla Reddy, A.Ch., and Lokesh, B.R. (1994) Effect of Dietary Turmerie (Curcuma longa) on Iron-Induced Lipid Peroxidation in the Rat Liver, Food Chem. Toxicol. 32, 279–283.

    Article  Google Scholar 

  6. 6.

    Elizabeth, K., and Rao, M.N.A. (1990) Oxygen Radical Scavenging Activity of Curcumin, Int. J. Pharm. 58, 237–240.

    Article  Google Scholar 

  7. 7.

    Unnikrishnan, M.K., and Rao, M.N.A. (1992) Curcumin Inhibits Nitrite Induced Methemoglobin Formation, FEBS Lett. 301, 195–196.

    PubMed  CAS  Article  Google Scholar 

  8. 8.

    Nagabhushan, M., Amonkar, A.J., and Bhide, S.U. (1987) In vitro Anti-mutagenecity of Curcumin Against Environmental Mutagens, Food Chem. Toxicol. 25, 545–548.

    PubMed  CAS  Article  Google Scholar 

  9. 9.

    Huang, M.-T., Wang, Z.Y., Georgiadis, C.A., Laskin, J.D., and Conney, A.H. (1991) Inhibitory Effects of Curcumin on Tumor Initiation by Benz[α]pyrene and 7,12-Dimethyl benz[α]anthracene, Carcinogenesis 13, 2183–2186.

    Google Scholar 

  10. 10.

    Li, C.J., Zhang, L.J., Dezube, B.J., Crumpacker, C.S., and Pardee, A. (1993) Three Inhibitors of Type I Human Immuno-deficiency Virus Long Terminal Repeat Directed Gene Expression and Virus Replications, Proc. Natl. Acad. Sci. USA 90, 1839–1842.

    PubMed  CAS  Article  Google Scholar 

  11. 11.

    Huang, M.-T., Smart, R.S., Wong, C.-Q., and Conney, A.H. (1988) Inhibitory Effect of Curcumin, Chlorogenic Acid, Caffeic Acid and Ferulic Acid on Tumor Promotion in Mouse Skin by12-O-Tetra-decanoylphorbol-13-acetate, Cancer. Res. 48, 5941–5946.

    PubMed  CAS  Google Scholar 

  12. 12.

    Huang, M.-T., Lysz, T., Ferraro, T., Abidi, T.F., Laskin, J.D., and Conney, A.H. (1991) Inhibitory Effects of Curcumin on in vitro Lipoxygenase and Cyclooxygenase Activities in Mouse Epidermis, Cancer Res. 51, 813–819.

    PubMed  CAS  Google Scholar 

  13. 13.

    Gorman, A.A., Hamblett, I., Hill, T.J., Jones, H., Srinivasan, V.S., and Wood, P.D. (1997) Gurcumin: A Pulse Radiolysis Investigation of the Radical in Micellar Systems, in Spices: Flavor Chemistry and Antioxidant Properties (Risch, S.J., and Howe, C.T., eds.), Vol. 660, pp. 235–243, American Chemical Society, Washington, DC.

    Google Scholar 

  14. 14.

    Kunchandy, E., and Rao, M.N.A. (1989) Effect of Curcumin on Hydroxyl Radical Generation Through Tocopherols and Tocotrienols, Int. J. Pharm. 57, 173–176.

    CAS  Article  Google Scholar 

  15. 15.

    Deodhar, S.D., Sethi, R., and Srimal, R.C. (1980) Preliminary Study on Anti-Rheumatic Activity of Curcumin (dirferuloyl methane), Ind. J. Med. Res. 701, 632–634.

    Google Scholar 

  16. 16.

    Srihari Rao, T., Basu, N., and Siddique, H.H. (1982) Anti-Inflammatory Activity of Curcumin Analogues, Ind. J. Med. Res. 75, 574–578.

    Google Scholar 

  17. 17.

    Ravindranath, V., and Chandrashekar, N. (1982) Metabolism of Curcumin: Studies with 3H-Curcumin, Toxicology 22, 337–344.

    CAS  Article  Google Scholar 

  18. 18.

    Spector, A.A. (1975) Fatty Acid Binding to Plasma Albumin, J. Lipid Res. 16, 165–179.

    PubMed  CAS  Google Scholar 

  19. 19.

    Tonnesen, H.H., and Greenhill, V.J. (1992) Studies on Curcumin and Curcuminiods: XXII: Curcumin as a Reducing Agent and as a Radical Scavenger, Int. J. Pharm. 87, 79–87.

    Article  Google Scholar 

  20. 20.

    Lehrer, S.S., and Fasman, J.D. (1966) The Fluorescence of Lysozyme and Lysozyme Substrate Complex, Biochem. Biophys. Res. Commun. 23, 133–138.

    PubMed  CAS  Article  Google Scholar 

  21. 21.

    Jaffe, H.H., and Orchin, M. (1962) in Theory and Applications of Ultraviolet Spectroscopy, John Wiley, New York, pp. 581–583.

    Google Scholar 

  22. 22.

    Ikeda, K., and Hamaguchi, K. (1969) The Binding of N-Acetyl-glucose Amine to Lysozyme; Studies on Circular Dichroism, J. Biochem. 66, 513–520.

    PubMed  CAS  Google Scholar 

  23. 23.

    Maliwal, B.P., Appu Rao, A.G., and Narasinga Rao, M.S. (1985) Spectroscopic Study of the Interaction of Gossypol with Bovine Serum Albumin, Int. J. Pep. Protein Res. 25, 382–388.

    CAS  Article  Google Scholar 

  24. 24.

    Oh, H.I., Hoff, J.E., Armstrong, G.S., and Haff, L.A. (1980) Hydrophobic Interaction in Tannin-Protein Complexes, J. Agric. Food Chem. 28, 394–398.

    CAS  Article  Google Scholar 

  25. 25.

    Haslam, E. (1996) Natural Polyphenols (vegetable tannins) as Drugs: Possible Modes of Action, J. Nat. Prod. 59, 205–215.

    PubMed  CAS  Article  Google Scholar 

  26. 26.

    Pederson, O.L., and Jacobson, M. (1980) Independent Binding of Ligands to Human Serum Albumin, J. Biol. Chem. 32, 5067–5072.

    Google Scholar 

  27. 27.

    Appu Rao, A.G. (1992) A Stoichiometric Analysis of Bovine Serum Albumin-Gossypol Interactions: A Fluorescence Quenching Study, Ind. J Biochem Biophys. 29, 179–182.

    Google Scholar 

  28. 28.

    Peters, T., Jr. (1985) Serum Albumin, Adv. Protein Chem. 37, 161–245.

    PubMed  CAS  Article  Google Scholar 

  29. 29.

    Jacobson, J., and Brodersen, R. (1983) Albumin-Bilirubin Binding Mechanism, J. Biol. Chem. 258, 6319–6326.

    Google Scholar 

  30. 30.

    Rechieri, G.V., Anel, A., and Kleinfeld, A.M. (1993) Interaction of Long-Chain Free Fatty Acids and Albumin: Determination of Free Fatty Acid Levels Using the Fluorescent Probe AD-IFAB, Biochemistry 32, 7574–7580.

    Article  Google Scholar 

  31. 31.

    Began, G., Sudharshan, E., and Appu Rao, A.G. (1998) Inhibition of Lipoxygense 1 by Phosphatidylcholine Micelles-Bound Curcumin, Lipids 33, 1223–1228.

    PubMed  CAS  Google Scholar 

  32. 32.

    Reddy, S., and Aggarwal, B.B. (1994) Cureumin Is a Competitive and Selective Inhibitor of Phosphorylase Kinase, FEBS Lett. 341, 19–22.

    PubMed  CAS  Article  Google Scholar 

  33. 33.

    Yamamoto, H., Hanada, K., Kawasaki, K., and Nishijima M. (1997) Inhibitory Effect of Curcumin on Mammalian Phospho-lipase D Activity. FEBS Lett. 417, 196–198.

    PubMed  CAS  Article  Google Scholar 

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Correspondence to A. G. Appu Rao.

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Pulla Reddy, A.C., Sudharshan, E., Appu Rao, A.G. et al. Interaction of curcumin with human serum albumin—A spectroscopic study. Lipids 34, 1025–1029 (1999). https://doi.org/10.1007/s11745-999-0453-x

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Keywords

  • Curcumin
  • Human Serum Albu
  • Circular Dichroism
  • Gossypol
  • Cumin