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Recombinant intracellular Rhodospirillum rubrum L-asparaginase with low L-glutaminase activity and antiproliferative effect

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Abstract

The recombinant producer strain expressing Rhodospirillum rubrum L-asparaginase (RrA) has been obtained and a purification procedure of RrA has been developed. The purified enzyme, RrA, has the following biochemical and catalytic characteristics: Km for L-Asn of 0.22 mM, pH optimum at 9.2; temperature optimum at 54°C, pI = 5.1. RrA exhibited a significant cytotoxic effect towards the following cell lines: K562 (IC50 = 1.80 U/mL), DU145 (IC50 = 9.19 U/mL), and MDA-MB-231 (IC50 = 34.62 U/mL). Comparative analysis employing E. coli L-asparaginase II type (EcA) and Erwinia carotovora L-asparaginase (EwA) has shown that the enzyme cytotoxicity towards these cell lines decreased in the following order: EcA > RrA > EwA. Daily administration of RrA (4000 U/kg) to L5178y bearing mice for 10 days (total dose of 40000 U/kg) showed T/C = 172. Data obtained suggest that RrA may be referred to intracellular L-asparaginases with low L-glutaminase activity and marked antiproliferative effect.

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References

  1. van den Berg, H., Leuk. Lymphoma, 2011, vol. 52, pp. 168–178.

    Article  Google Scholar 

  2. Jaccard, A., Petit, B., Girault, S., Suarez, F., Gressin, R., Zini, J.M., Coiteux, V., Larroche, C., Devidas, A., Thiéblemont, C., Gaulard, P., Marin, B., Gachard, N., Bordessoule, D., and Hermine, O., Ann. Oncol., 2009, vol. 20, pp. 110–116.

    Article  CAS  Google Scholar 

  3. Yong, W., Zheng, W., Zhang, Y., Zhu, J., Wei, Y., Zhu, D., and Li, J., Int. J. Hematol., 2003, vol. 78, pp. 163–167.

    Article  CAS  Google Scholar 

  4. Obama, K., Tara, M., and Niina, K., Int. J. Hematol., 1999, vol. 69, pp. 260–262.

    CAS  Google Scholar 

  5. Agrawal, N.R., Bukowski, R.M., Rybicki, L.A., Kurtzberg, J., Cohen, L.J., and Hussein, M.A., Cancer, 2003, vol. 98, pp. 94–99.

    Article  CAS  Google Scholar 

  6. Kobrinsky, N.L., Sposto, R., Shah, N.R., Anderson, J.R., DeLaat, C., Morse, M., Warkentin, P., Gilchrist, G.S., Cohen, M.D., Shina, D., and Meadows, A.T., J. Clin. Oncol., 2001, vol. 19, pp. 2390–2396.

    CAS  Google Scholar 

  7. Reinert, R.B., Oberle, L.M., Wek, S.A., Bunpo, P., Wang, X.P., Mileva, I., Goodwin, L.O., Aldrich, C.J., Durden, D.L., McNurlan, M.A., Wek, R.C., and Anthony, T.G., J. Biol. Chem., 2006, vol. 281, pp. 31222–31233.

    Article  CAS  Google Scholar 

  8. Ollenschläger, G., Roth, E., Linkesch, W., Jansen, S., Simmel, A., and Mödder, B., Eur. J. Clin. Invest., 1988, vol. 18, pp. 512–516.

    Article  Google Scholar 

  9. Villa, P., Corada, M., and Bartosek, I., Toxicol. Lett., 1986, vol. 32, pp. 235–241.

    Article  CAS  Google Scholar 

  10. Warrell, R.P. Jr., Chou, T.C., Gordon, C., Tan, C., Roberts, J., Sternberg, S.S., Philips, F.S., and Young, C.W., Cancer Res., 1980, vol. 40, pp. 4546–4551.

    Google Scholar 

  11. Capizzy, R.L. and Cheng, Y.C., in Enzymes as Drugs, Holcenberg, J.S., Roberts, J., Eds., New York: John Wiley and Sons, 1981, pp. 1–24.

    Google Scholar 

  12. Storti, E. and Quaglino, D., in: Experimental and Clinical Effects of L-Asparaginase, Grundmann, E., and Oettgen, H.F., Eds., Berlin-Heidelberg-New York: Springer-Verlag, 1970, pp. 344–349.

    Chapter  Google Scholar 

  13. Bendich, A., Kafkewitz, D., Abuchowski, A., and Davis, F.F., Clin. Exp. Immunol., 1982, vol. 48, pp. 273–278.

    CAS  Google Scholar 

  14. Distasio, J.A., Salazar, A.M., Nadji, M., and Durden, D.L., Int. J. Cancer, 1982, vol. 30, pp. 343–347.

    Article  CAS  Google Scholar 

  15. Gladilina, Yu.A., Sokolov, N.N., and Krasotkina, Yu.V., Biomed. Khim., 2008, vol. 54, pp. 482–486.

    CAS  Google Scholar 

  16. Cappelletty, D., Chiarelli, L.R., Pasquetto, M.V., Stivala, S., Valentini, G., and Scotti, C., Biochem. Biophys. Res. Commun., 2008, vol. 377, pp. 1222–1226.

    Article  Google Scholar 

  17. Ghosh, R., Hardmeyer, A., Thoenen, I., and Bachofen, R., Appl. Environ. Microbiol., 1994, vol. 60, pp. 1698–1700.

    CAS  Google Scholar 

  18. Neilson, A.H. and Nordlund, S., J. Gen. Microbiol., 1975, vol. 91, pp. 53–62.

    CAS  Google Scholar 

  19. Sambrook, J. and Russell, D.W., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor, 2001.

    Google Scholar 

  20. Pokrovskaya, M.V., Pokrovsky, V.S., and Sokolov, N.N., Prikl. Biokhim. Mikrobiol., 2011, vol. 47, pp. 183–186.

    Google Scholar 

  21. Hartree, E.F., Anal. Biochem., 1991, vol. 192, pp. 215–218.

    Article  Google Scholar 

  22. Osterman, L.A., Issledovanie biologicheskikh makromolekul elektrofokusirovaniem, immunoelektroforezom i radioisotopnymi metodami (Study of Biological Macro-molecules by Electrofocusing, Immune Electrophoresis and Radioisotope Methods), Moscow: Nauka, 1983.

    Google Scholar 

  23. Borisova, A.A., Eldarov, M.A., Zhgun, A.A, Alexandrova, S.S., Omelyanyuk, N.M., Sokov, B.N., and Sokolov, N.N., Biomed. Khim., 2003, vol. 49, pp. 502–507.

    CAS  Google Scholar 

  24. Wade, H.E. and Philips, B.P., Anal. Biochem., 1971, vol. 72, p. 248.

  25. Wriston, J.C. and Yellin, T.O., Adv. Enzymol., 1985, vol. 39, pp. 185–248.

    Google Scholar 

  26. Dawson, R., Elliott, D., Elliott, W., and Jones, K.M., Spravochnik biokhimika (Data for Biochemical Research), Moscow: Mir, 1991.

    Google Scholar 

  27. Mossman, T., J. Immunol. Methods, 1983, vol. 65, pp. 55–63.

    Article  Google Scholar 

  28. Treshchalina, E.M., Protivoopukholevaya activnost’ veshchestv prirodnogo proiskhozhdeniya (The Antitumor Activity of Natural Substances), Moscow: Prakticheskaya Meditsina, 2005.

    Google Scholar 

  29. Chabner, B.A. and Longo, D.L., Cancer Chemotherapy and Biotherapy: Principles and Practice, Philadelphia: Lippincott-Raven, 2001.

    Google Scholar 

  30. Peng, G., Fritzsch, G., Zickermann, V. Schägger, H., Mentele, R., Lottspeich, F., Bostina, M., Radermacher, M., Huber, R., Stetter, K.O., and Michel, H., Biochemistry, 2003, vol. 42, pp. 3032–3039.

    Article  CAS  Google Scholar 

  31. Derst, C., Henseling, J., and Röhm, K.H., Protein Science, 2000, vol. 9, pp. 2009–2017.

    Article  CAS  Google Scholar 

  32. Michalska, K. and Jaskolski, M., Acta Biochim. Pol., 2006, vol. 53, pp. 627–640.

    CAS  Google Scholar 

  33. Bonthron, D.T. and Jaskolski, M., Acta Biochim. Pol., 1997, vol. 44, pp. 491–504.

    CAS  Google Scholar 

  34. Yao, M., Yasutake, Y., Morita, H., and Tanaka, I., Acta Crystallogr., 2005, vol. 61, pp. 294–301.

    Google Scholar 

  35. Campbell, H.A., Mashburn, L.T., Boyse, E.A., and Old, L.J., Biochemistry, 1967, vol. 6, pp. 721–730.

    Article  CAS  Google Scholar 

  36. Schwartz, J.H., Reeves, J.Y., and Broome, J.D., Proc. Natl. Acad. Sci. USA, 1966, vol. 56, pp. 1516–1519.

    Article  CAS  Google Scholar 

  37. Yun, M.K., Nourse, A., White, S.W., Rock, C.O., and Heath, R.J., J. Mol. Biol., 2007, vol. 369, pp. 794–811.

    Article  CAS  Google Scholar 

  38. Kitto, G.B., Smith, G., Thiet, T., Mason, M., and Davidson, L., J. Bacteriol., 1979, vol. 137, pp. 204–212.

    CAS  Google Scholar 

  39. Davidson, L., Brear, D.R., Wingard, P., Hawkins, J., and Kitto, G.B., J. Bacteriol., 1977, vol. 129, pp. 1379–1386.

    CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Biomedical Chemistry of Russian Academy of Medical Sciences, ul. Pogodinskaya 10, Moscow, 119121, Russia

    M. V. Pokrovskaya, V. S. Pokrovskiy, S. S. Aleksandrova, G. V. Ponomarev & N. N. Sokolov

  2. Blokhin Cancer Research Center of Russian Academy of Medical Sciences, Kashirskoye shosse 24, Moscow, 115478, Russia

    V. S. Pokrovskiy, N. Yu. Anisimova & E. M. Treschalina

  3. Bach Institute of Biochemistry of Russian Academy of Sciences, Leninsky prospekt 33, bld. 2, Moscow, 119071, Russia

    R. M. Andrianov

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  1. M. V. Pokrovskaya
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  2. V. S. Pokrovskiy
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  3. S. S. Aleksandrova
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  4. N. Yu. Anisimova
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  5. R. M. Andrianov
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  6. E. M. Treschalina
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  7. G. V. Ponomarev
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  8. N. N. Sokolov
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Correspondence to V. S. Pokrovskiy.

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Original Russian Text © M.V. Pokrovskaya, V.S. Pokrovskiy, S.S. Aleksandrova, N.Yu. Anisimova, R.M. Andrianov, E.M. Treschalina, G.V. Ponomarev, N.N. Sokolov, 2012, published in Biomeditsinskaya Khimiya.

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Pokrovskaya, M.V., Pokrovskiy, V.S., Aleksandrova, S.S. et al. Recombinant intracellular Rhodospirillum rubrum L-asparaginase with low L-glutaminase activity and antiproliferative effect. Biochem. Moscow Suppl. Ser. B 6, 123–131 (2012). https://doi.org/10.1134/S1990750812020096

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  • DOI: https://doi.org/10.1134/S1990750812020096

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