Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Construction of Novel Bifunctional Chimeric Proteins Possessing Antitumor and Thrombolytic Activities

  • Hui, Jing (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University) ;
  • Dai, Youjin (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University) ;
  • Bian, Yuanyuan (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University) ;
  • Li, Hui (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University) ;
  • Cui, Xiaojin (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University) ;
  • Yu, Xiaojie (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University) ;
  • You, Song (College of Pharmaceutical Engineering, Shenyang Pharmaceutical University) ;
  • Hu, Fengqing (Laboratory of Biomaterials and Biopharmaceuticals, School of Life Sciences, Liaoning University)
  • Received : 2011.07.26
  • Accepted : 2012.03.07
  • Published : 2012.07.28
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Abstract

Based on their respective antitumor and thrombolytic activities, the superantigen staphylococcal enterotoxin C2 (SEC2) and staphylokinase (Sak) were chosen for the construction of the novel chimeric proteins Sak-linker-SEC2 and SEC2-linker-Sak using a linker composed of nine Ala residues. Both chimeric proteins possessed nearly the same PBMC proliferation stimulating activity and antitumor activity as SEC2 and thrombolytic activity as Sak. Neither the SEC2 or Sak component of each chimeric protein affected the activity of the other component. The results presented in this study provide a possible strategy to prevent and cure tumor thrombus.

Keywords

References

  1. Abrahmsen, L. 1995. Superantigen engineering. Curr. Opin. Struct. Biol. 5: 464-470. https://doi.org/10.1016/0959-440X(95)80030-1
  2. Alpaugh, R. K., J. Schultz, C. McAleer, B. J. Giantonio, R. Persson, M. Burnite, et al. 1998. Superantigen-targeted therapy: Phase I escalating repeat dose trial of the fusion protein PNU-214565 in patients with advanced gastrointestinal malignancies. Clin. Cancer Res. 4: 1903-1914.
  3. Andreescu, A. C. M., M. Cushman, J. M. Hammond, and M. E. Wood. 2001. Trousseau's syndrome treated with long-term subcutaneous lepirudin (case report and review of the literature). J. Thromb. Thrombolys. 11: 33-37. https://doi.org/10.1023/A:1008952126124
  4. Baker, M. D. and K. R. Acharya. 2004. Superantigens: Structure-function relationships. Int. J. Med. Microbiol. 293: 529-537. https://doi.org/10.1078/1438-4221-00298
  5. Bokarewa, M. I., T. Jin, and A. Tarkowski. 2006. Staphylococcus aureus: Staphylokinase. Int. J. Biochem. Cell Biol. 38: 504-509 https://doi.org/10.1016/j.biocel.2005.07.005
  6. Brener, S. J. and E. J. Topol. 1998. Third-generation Thrombolytic Agents for Acute Myocardial Infarction. Marcel Dekker Inc., New York.
  7. Chen, T. Z. 2001. The exploitation of HAS and its application in tumor therapy. Prog. Microbiol. Immunol. 29: 63-69.
  8. Chen, T. Z. 2005. Gaojusheng: A novel anti-cancer drug prepared from SEC superantigen. Prog. Microbiol. Immunol. 33: 49-50.
  9. Chew, H. K., T. Wun, D. Harvey, H. Zhou, and R. H. White. 2006. Incidence of venous thromboembolism and its effect on survival among patients with common cancers. Arch. Intern. Med. 166: 458-464.
  10. Collen, D., B. Schlott, Y. Engelborghs, B. Van Hoet, M. Hartmann, H. R. Lijnen, and D. Behnke. 1993. On the mechanism of the activation of human plasminogen by recombinant staphylokinase. J. Biol. Chem. 268: 8284-8289.
  11. Dohlsten, M., A. Sundstedt, M. Bjorklund, G. Hedlund, and J. Kalland. 1993. Superantigen-induced cytokines suppress growth of human colon-carcinoma cells. Int. J. Cancer 54: 482-488. https://doi.org/10.1002/ijc.2910540321
  12. Grossman, D., J. G. Lamphear, J. A. Mollick, M. J. Betley, and R. R. Rich. 1992. Dual roles for class II major histocompatibility complex molecules in staphylococcal enterotoxin-induced cytokine production and in vivo toxicity. Infect. Immun. 60: 5190-5196.
  13. Hansson, J., L. Ohlsson, R. Persson, G. Andersson, N. G. Ilbäck, M. J. Litton, et al. 1997. Genetically engineered superantigens as tolerable antitumor agents. Proc. Natl. Acad. Sci. USA 94: 2489-2494. https://doi.org/10.1073/pnas.94.6.2489
  14. Hedlund, G., M. Dohlsten, C. Petersson, and T. Kalland. 1993. Superantigen-based tumor therapy: In vivo activation of cytotoxic T cells. Cancer Immunol. Immunother. 36: 89-93. https://doi.org/10.1007/BF01754407
  15. Herman, A., J. W. Kappler, P. Marrack, and A. M. Pullen. 1991. Superantigens: Mechanism of T-cell stimulation and role in immune responses. Annu. Rev. Immunol. 9: 745-772. https://doi.org/10.1146/annurev.iy.09.040191.003525
  16. Hui, J., Y. Cao, F. Xiao, J. Zhang, H. Li, and F. Q. Hu. 2008. Staphylococcus aureus enterotoxin C2 mutants: Biological activity assay in vitro. J. Ind. Microbiol. Biotechnol. 35: 975-980 https://doi.org/10.1007/s10295-008-0372-3
  17. Johnson, H. M., J. K. Russell, and C. H. Pontzer. 1991. Staphylococcal enterotoxin microbial superantigens. FASEB J. 5: 2706-2712.
  18. Kakkar, A. K. 2009. Antithrombotic therapy and survival in cancer patients. Best Pract. Res. Clin. Haematol. 22: 147-151. https://doi.org/10.1016/j.beha.2009.01.004
  19. Kwaan, H. C. and B. Vicuna. 2007. Thrombosis and bleeding in cancer patients. Oncol. Rev. 1: 14-27. https://doi.org/10.1007/s12156-007-0003-7
  20. Lip, G. Y., B. S. Chin, and A. D. Blann. 2002. Cancer and the prothrombotic state. Lancet Oncol. 3: 27-34. https://doi.org/10.1016/S1470-2045(01)00619-2
  21. Ma, W. X., H. Yu, Q. Q. Wang, J. F. Bao, J. Yan and H. Jin. 2004. In vitro biological activities of transmembrane superantigen staphylococcal enterotoxin A fusion protein. Cancer Immunol. Immuother. 53: 118-124. https://doi.org/10.1007/s00262-003-0437-0
  22. Mandi, N., S. Soorapaneni, S. Rewanwar, P. Kotwal, B. Prasad, G. Mandal, and S. Padmanabhan. 2009. High yielding recombinant staphylokinase in bacterial expression, purification and activity studies. Protein Expr. Purif. 64: 69-75. https://doi.org/10.1016/j.pep.2008.10.010
  23. Matsumoto, K., K. Takase, Y. Yamamoto, Y. Doi, and S. Taguchi. 2009. Chimeric enzyme composed of polyhydroxyalkanoate (PHA) synthases from Ralstonia eutropha and Aeromonas caviae enhances production of PHAs in recombinant Escherichia coli. Biomacromolecules 10: 682-685. https://doi.org/10.1021/bm801386j
  24. Mondal, T. K., D. Bhatta, S. Biswas, and P. Pal. 2002. Superantigen-induced apoptotic death of tumor cells is mediated by cytotoxic lymphocytes, cytokines, and nitric oxide. Biochem. Biophys. Res. Commun. 290: 1336-1342. https://doi.org/10.1006/bbrc.2002.6359
  25. Ochi, A., K. Migita, J. Xu, and K. Siminovitch. 1993. In vivo tumor immunotherapy by a bacterial superantigen. J. Immunol. 151: 3180-3186.
  26. Pardoll, D. M. 1995. Paracrine cytokine adjuvants in cancer immunotherapy. Annu. Rev. Immunol. 13: 399-415. https://doi.org/10.1146/annurev.iy.13.040195.002151
  27. Rickles, F. R. and M. N. Levine. 2001. Epidemiology of thrombosis in cancer. Acta Haematol. 106: 6-12. https://doi.org/10.1159/000046583
  28. Saksela, O. 1981. Radial caseinolysis in agarose: A simple method for detection of plasminogen activator in the presence of inhibitory substances and serum. Anal. Biochem. 111: 276-282 https://doi.org/10.1016/0003-2697(81)90564-9
  29. Sambrook, J. and D. W. Russell. 2001. Molecular Cloning: A Laboratory Manual, 3rd Ed. Cold Spring Harbor Laboratory Press, New York.
  30. Scholl, P. R., N. Trede, T. A. Chatila, and R. S. Geha. 1992. Role of protein tyrosine phosphorylation in monokine induction by the staphylococcal superantigen toxic shock syndrome toxin-1. J. Immunol. 148: 2237-2241.
  31. Sundstedt, A., M. Celander, and G. Hedlund. 2008. Combining tumor-targeted superantigens with interferon-alpha results in synergistic anti-tumor effects. Int. Immunopharmacol. 8: 442-452 https://doi.org/10.1016/j.intimp.2007.11.006
  32. Toombs, C. F. 2001. New directions in thrombolytic therapy. Curr. Opin. Pharmacol. 1: 164-168. https://doi.org/10.1016/S1471-4892(01)00030-3
  33. Verstraete, M. 2000. Third-generation thrombolytic drugs. Am. J. Med. 109: 52-58. https://doi.org/10.1016/S0002-9343(00)00380-6
  34. Wang, J. Z. and M. Fan. 2002. Manual of Protein Technology. Science Press, Beijing.
  35. Xu, M. K. and C. G. Zhang. 2006. Gene expression and function study of fusion immunotoxin anti-Her-2-scFv-SEC2 in Escherichia coli. Appl. Microbiol. Biotechnol. 70: 78-84. https://doi.org/10.1007/s00253-005-0049-z

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