Journal of Fluorescence

, Volume 29, Issue 4, pp 827–835 | Cite as

Probing the Interaction of Newly Synthesized Pt(II) Complex on Human Serum Albumin Using Competitive Binding Site Markers

  • Akram Najaran
  • Adeleh DivsalarEmail author
  • Ali Akbar Saboury
  • Nasim Hayati Roodbari


Considering the importance of pharmacology and the influence of drugs on biological materials, the effects of a newly designed and synthesized platin complex (2,2′-Bipyridine-3,3′-dicarboxylic acid, oxalato Platinum(II), as an antitumor drug was tested on the structure of blood carrier protein of human serum albumin (HSA) using various spectroscopic techniques including UV-visible, fluorescence, and circular dichroism at 25 and 37 °C. Results of the fluorescence measurements revealed that adding the complex caused reduction in intrinsic fluorescence emission of HSA resulted from dynamic quenching of HSA. The number of binding sites and binding constants were calculated at both temperatures of 25 and 37 °C. In addition, in order to identify the complex’s binding site on HSA employing spectroscopy, the competitive studies were followed using warfarin, digitoxin and ibuprofen as site markers of Sudlow sites I, II and III. Competitive binding test results have shown that Pt(II) complex bind on the warfarin binding site (or Sudlow sites I) on HSA. Besides, a reduction in thermal stability for HSA was observed in the presence of the newly designed Pt(II) complex.


Site markers HSA Sudlow sites Competitive studies Fluorescence quenching 



Authors thank the Research Council of Kharazmi University for financial support of this work.


  1. 1.
    Varshney A, Sen P, Ahmad E, Rehan M, Subbarao N, Khan RH (2010) Ligand binding strategies of human serum albumin: how can the cargo be utilized? Chirality 22(1):77–87CrossRefGoogle Scholar
  2. 2.
    Keppler BK (1993) Metal complexes in cancer chemotherapy. Wiley-VCH, Weinheim & New YorkGoogle Scholar
  3. 3.
    Kragh-Hansen U, Watanabe H, Nakajou K, Iwao Y, Otagiri M (2006) Chain length-dependent binding of fatty acid anions to human serum albumin studied by site-directed mutagenesis. J Mol Biol 363(3):702–712CrossRefGoogle Scholar
  4. 4.
    Divsalar A, Bagheri MJ, Saboury AA, Mansoori-Torshizi H, Amani M (2009) Investigation on the interaction of newly designed anticancer Pd (II) complexes with different aliphatic tails and human serum albumin. J Phys Chem B 113(42):14035–14042CrossRefGoogle Scholar
  5. 5.
    Yang F, Bian C, Zhu L, Zhao G, Huang Z, Huang M (2007) Effect of human serum albumin on drug metabolism: structural evidence of esterase activity of human serum albumin. J Struct Biol 157(2):348–355CrossRefGoogle Scholar
  6. 6.
    Zunszain PA, Ghuman J, Komatsu T, Tsuchida E, Curry S (2003) Crystal structural analysis of human serum albumin complexed with hemin and fatty acid. BMC Struct Biol 3(1):6CrossRefGoogle Scholar
  7. 7.
    Yamasaki K, Maruyama T, Kragh-Hansen U, Otagiri M (1996) Characterization of site I on human serum albumin: concept about the structure of a drug binding site. Biochim Biophys Acta 1295(2):147–157CrossRefGoogle Scholar
  8. 8.
    Ascenzi P, Fasano M (2010) Allostery in a monomeric protein: the case of human serum albumin. Biophys Chem 148(1–3):16–22CrossRefGoogle Scholar
  9. 9.
    Espósito BP, Najjar R (2002) Interactions of antitumoral platinum-group metallodrugs with albumin. Coord Chem Rev 232(1–2):137–149CrossRefGoogle Scholar
  10. 10.
    Curry S, Mandelkow H, Brick P, Franks N (1998) Crystal structure of human serum albumin complexed with fatty acid reveals an asymmetric distribution of binding sites. Nat Struct Mol Biol 5(9):827–835CrossRefGoogle Scholar
  11. 11.
    Petitpas I, Bhattacharya AA, Twine S, East M, Curry S (2001) Crystal structure analysis of warfarin binding to human serum albumin anatomy of drug site I. J Biol Chem 276(25):22804–22809CrossRefGoogle Scholar
  12. 12.
    Sudlow G, Birkett D, Wade D (1976) Further characterization of specific drug binding sites on human serum albumin. Mol Pharmacol 12(6):1052–1061PubMedGoogle Scholar
  13. 13.
    Liu X, Li S, Zhang J, Chen X (2009) Flow injection-capillary electrophoresis frontal analysis method for the study of the interactions of a series of drugs with human serum albumin. J Chromatogr B 877(27):3144–3150CrossRefGoogle Scholar
  14. 14.
    Divsalar A, Khodabakhshian S (2015) Probing the binding site of a new synthesized anti-cancer compound to HSA via competitive ligand binding method. J Mol Liq 206:82–88CrossRefGoogle Scholar
  15. 15.
    DESOIZE B (2004) Metals and metal compounds in cancer treatment. Anticancer Res 24(3A):1529–1544PubMedGoogle Scholar
  16. 16.
    Tušek-Božić L, Juribašić M, Traldi P, Scarcia V, Furlani A (2008) Synthesis, characterization and antitumor activity of palladium (II) complexes of monoethyl 8-quinolylmethylphosphonate. Polyhedron 27(4):1317–1328CrossRefGoogle Scholar
  17. 17.
    Polyanskaya TV, Kazhdan I, Motley DM, Walmsley JA (2010) Synthesis, characterization and cytotoxicity studies of palladium (II)–proflavine complexes. J Inorg Biochem 104(11):1205–1213CrossRefGoogle Scholar
  18. 18.
    Pourgonabadi S, Reza Saberi M, Khan Chamani J (2011) Investigating the antagonistic action between aspirin and tamoxifen with hsa: identification of binding sites in binary and ternary drug-protein systems by spectroscopic and molecular modeling approaches. Protein Pept Lett 18(3):305–317CrossRefGoogle Scholar
  19. 19.
    Ghalandari B, Divsalar A, Saboury AA, Haertlé T, Parivar K, Bazl R, Eslami-Moghadam M, Amanlou M (2014) Spectroscopic and theoretical investigation of oxali-palladium interactions with β-lactoglobulin. Spectrochim Acta A Mol Biomol Spectrosc 118:1038–1046CrossRefGoogle Scholar
  20. 20.
    Saeidifar M, Mansouri-Torshizi H, Divsalar A, Saboury AA (2013) Spectroscopic investigation on the binding of the antitumoral Pd (II) complex to human serum albumin. J Chin Chem Soc 60(2):133–139CrossRefGoogle Scholar
  21. 21.
    Lakowicz JR (2006) Principles of fluorescence spectroscopy, 3rd edn. Springer, New YorkCrossRefGoogle Scholar
  22. 22.
    Abbasi-Tajarag K, Divsalar A, Saboury AA, Ghalandari B, Ghourchian H (2015) Destructive effect of anticancer oxali-palladium on heme degradation through the generation of endogenous hydrogen peroxide. J Biomol Struct Dyn 34(11):2493–2504CrossRefGoogle Scholar
  23. 23.
    Song M, Liu S, Yin J, Wang H (2011) Interaction of human serum album and C60 aggregates in solution. Int J Mol Sci 12(8):4964–4974CrossRefGoogle Scholar
  24. 24.
    Curry S (2002) Beyond expansion: structural studies on the transport roles of human serum albumin. Vox Sang 83:315–319CrossRefGoogle Scholar
  25. 25.
    Divsalar A, Saboury AA, Mansoori-Torshizi H, Ahmad F (2010) Design, synthesis, and biological evaluation of a new palladium (II) complex: β-lactoglobulin and K562 as targets. J Phys Chem B 114(10):3639–3647CrossRefGoogle Scholar
  26. 26.
    Kratochwil NA, Huber W, Müller F, Kansy M, Gerber PR (2002) Predicting plasma protein binding of drugs: a new approach. Biochem Pharmacol 64(9):1355–1374CrossRefGoogle Scholar
  27. 27.
    Ross PD, Subramanian S (1981) Thermodynamics of protein association reactions: forces contributing to stability. Biochemistry 20(11):3096–3102CrossRefGoogle Scholar
  28. 28.
    Mallick A, Haldar B, Chattopadhyay N (2005) Spectroscopic investigation on the interaction of ICT probe 3-acetyl-4-oxo-6, 7-dihydro-12H indolo-[2, 3-a] quinolizine with serum albumins. J Phys Chem B 109(30):14683–14690CrossRefGoogle Scholar
  29. 29.
    Yue Y, Chen X, Qin J, Yao X (2009) Spectroscopic investigation on the binding of antineoplastic drug oxaliplatin to human serum albumin and molecular modeling. Colloids Surf B: Biointerfaces 69(1):51–57CrossRefGoogle Scholar
  30. 30.
    Divsalar A, Saboury AA, Ahadi L, Zemanatiyar E, Mansouri-Torshizi H (2010) Investigation of effects of newly synthesized Pt (II) complex against human serum albumin and leukemia cell line of K562. BMB Rep 43(11):766–771CrossRefGoogle Scholar
  31. 31.
    Singha Roy A, Ghosh KS, Dasgupta S (2013) An investigation into the altered binding mode of green tea polyphenols with human serum albumin on complexation with copper. J Biomol Struct Dyn 31(10):1191–1206CrossRefGoogle Scholar
  32. 32.
    Curry S (2009) Lessons from the crystallographic analysis of small molecule binding to human serum albumin. Drug Metab Pharmacokinet 24(4):342–357CrossRefGoogle Scholar
  33. 33.
    Sarraf N et al (2005) Thermodynamic studies on the interaction of copper ions with carbonic anhydrase. Bull Kor Chem Soc 26(7):1051–1056CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Akram Najaran
    • 1
  • Adeleh Divsalar
    • 2
    Email author
  • Ali Akbar Saboury
    • 3
  • Nasim Hayati Roodbari
    • 1
  1. 1.Department of Biology, Science and Research BranchIslamic Azad UniversityTehranIran
  2. 2.Department of Cell & Molecular Sciences, Faculty of Biological SciencesKharazmi UniversityTehranIran
  3. 3.Institute of Biochemistry and BiophysicsUniversity of TehranTehranIran

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