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In vitro production of three-finger neurotoxins from snake venoms, a disulfide rich proteins. Problems and their solutions (Review)

Russian Journal of Bioorganic Chemistry volume 36, pages 137–145 (2010)Cite this article

On 50-year anniversary of the Institute of Bioorganic Chemistry

Abstract

α-Neurotoxins from snake venom are highly efficient inhibitors of nicotinic acetylcholine receptors (nAChRs). These small proteins with a β-structural organization attract much interest as a tool for studies of nAChR and as prototypes for the development of new pharmaceuticals for the treatment of diseases of the nervous system. However, in vitro production of “three-finger” neurotoxins is complicated by the presence of four or five spatially closed disulfide bonds in their molecules. The present review contains a description of the most frequently used modern approaches for the E. coli expression of recombinant proteins (direct expression, expression as hybrid proteins, and secretion) with an emphasis placed on the recombinant production of snake α-neurotoxins. The methods of E. coli expression of isotopically labeled neurotoxins are described. The proposed solutions will be of broad interest for the bacterial production of other disulfide rich proteins.

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Abbreviations

GSH and GSSH:

reduced and oxidized forms of glutathione

GST:

glutathione-transferase

IC50 :

inhibition constant, at wich 50% of binding sites of receptor is blocked

nAChR:

nicotine acetylcholine receptor

NTII:

neurotoxin of venom of Naja oxiana

NTII/NTI:

chimeric toxin obtained by introduction onto the end of central loop of neurotoxin II of a fragment of the central loop of neurotoxin I of venom of Naja oxiana with additional disulfide bond

STII:

signal peptide of enterotoxin II of E. coli

TRX:

thioredoxine

WTX:

“weak” toxin of venom of Naja kaouthia

ZZ:

domen—synthetic IgG-binding ZZ-domen of protein A of staphillococci

References

  1. Tsetlin, V.I. and Hucho, F., FEBS Lett., 2004, vol. 557, nos. 1–3, pp. 9–13.

    Article  CAS  PubMed  Google Scholar 

  2. D’hoedt, D. and Bertrand, D., Expert Opin. Ther. TarGets, 2009, vol. 13, no. 4, pp. 395–411.

    Article  PubMed  Google Scholar 

  3. Hogg, R.C., Raggenbass, M., and Bertrand, D., Rev. Physiol. Biochem. Pharmacol., 2003, vol. 147, pp. 1–46.

    Article  CAS  PubMed  Google Scholar 

  4. Bourne, Y., Talley, T.T., Hansen, S.B., Taylor, P., and Marchot, P., EMBO J., 2005, vol. 24, pp. 1512–1522.

    Article  CAS  PubMed  Google Scholar 

  5. Nirthanan, S. and Gwee, M.C., J. Pharmacol. Sci., 2004, vol. 94, no. 1, pp. 1–17.

    Article  CAS  PubMed  Google Scholar 

  6. Golovanov, A.P., Lomize, A.L., Arseniev, A.S., Utkin, Y.N., and Tsetlin, V.I., Eur. J. Biochem., 1993, vol. 213, pp. 1213–1223.

    Article  CAS  PubMed  Google Scholar 

  7. Walkinshaw, M.D., Saenger, W., and Maelicke, A., Proc. Natl. Acad. Sci. USA, 1980, vol. 77, pp. 2400–2404.

    Article  CAS  PubMed  Google Scholar 

  8. Mordvintsev, D.Y., Polyak, Y.L., Kuzmin, D.A., Levtsova, O.V., Tourleigh, Y.V., Utkin, Y.N., Shaitan, K.V., and Tsetlin, V.I., Comput. Biol. Chem., 2007, vol. 31, no. 2, pp. 72–81.

    Article  CAS  Google Scholar 

  9. Khechinashvili, N.N. and Tsetlin, V.I., Mol. Biol. (Moscow), 1984, vol. 18, no. 3, pp. 786–791.

    CAS  Google Scholar 

  10. Gilquin, B., Bourgoin, M., Ménez, R., Le Du, M.H., Servent, D., Zinn-Justin, S., and Ménez, A., Protein. Sci., 2003, vol. 12, no. 2, pp. 266–277.

    Article  CAS  PubMed  Google Scholar 

  11. Ploug, M. and Ellis, V., FEBS Lett., 1994, vol. 349, no. 2, pp. 163–168.

    Article  CAS  PubMed  Google Scholar 

  12. Miwa, J.M., Ibanez-Tallon, I., Crabtree, G.W., Sanchez, R., Sali, A., Role, L.W., and Heintz, N., Neuron, 1999, vol. 23, no. 1, pp. 105–114.

    Article  CAS  PubMed  Google Scholar 

  13. Dessaud, E., Salaun, D., Gayet, O., and Chabbert, M., Mol. Cell Neurosci., 2006, vol. 31, no. 2, pp. 232–242.

    Article  CAS  PubMed  Google Scholar 

  14. Adermann, K., Wattler, F., Wattler, S., Heine, G., Meyer, M., Forssmann, W.G., and Nehls, M., Protein Sci., 1999, vol. 8, no. 4, pp. 810–819.

    CAS  PubMed  Google Scholar 

  15. Tsuji, H., Okamoto, K., Matsuzaka, Y., Iizuka, H., Tamiya, G., and Inoko, H., Genomics, 2003, vol. 81, no. 1, pp. 26–33.

    Article  CAS  PubMed  Google Scholar 

  16. Miwa, M., Stevens, T.R., King, S.L., Caldarone, B.J., Ibanez-Tallon, I., Xiao, C., Fitzsimonds, R.M., Pavlides, C., Lester, H.A., Picciotto, M.R., and Heintz, N., Neuron, 2006, vol. 51, no. 5, pp. 587–600.

    Article  CAS  PubMed  Google Scholar 

  17. Sørensen, H.P. and Mortensen, K.K., J. Biotechnol., 2005, vol. 115, no. 2, pp. 113–128.

    Article  PubMed  CAS  Google Scholar 

  18. Berndt, C., Lillig, C.H., and Holmgren, A., Biochim. Biophys. Acta, 2008, vol. 1783, no. 4, pp. 641–650.

    Article  CAS  PubMed  Google Scholar 

  19. McCoy, J. and Lavallie, E., Curr. Protoc. Mol. Biol., 2001, Ch. 16: Unit 16.8.

  20. Sheehan, D., Meade, G., Foley, V.M., and Dowd, C.A., Biochem. J., 2001, vol. 360, Pt. 1, pp. 1–16.

    Article  CAS  PubMed  Google Scholar 

  21. Maina, C.V., Riggs, P.D., and Grandea, A.G., 3rd, Slatko B.E., Moran, L.S., Tagliamonte, J.A., McReynolds, L.A., and Guan, C.D., Gene, 1988, vol. 74, no. 2, pp. 365–373.

    Article  CAS  PubMed  Google Scholar 

  22. Inouye, S. and Sahara, Y., Protein Expr. Purif., 2009, vol. 66,no. 1, pp. 52–57.

    Article  CAS  PubMed  Google Scholar 

  23. Marston, F.A., Biochem. J., 1986, vol. 240, no. 1, pp. 1–12.

    CAS  PubMed  Google Scholar 

  24. LaVallie, E.R., DiBlasio, E.A., Kovacic, S., Grant, K.L., Schendel, P.F., and McCoy, J.M., Biotechnology (N. Y.), 1993, vol. 11,no. 2, pp. 187–193.

    Article  CAS  Google Scholar 

  25. Kapust, R.B., Tözsér, J., Copeland, T.D., and Waugh, D.S., Biochem. Biophys. Res. Commun., 2002, vol. 294, no. 5, pp. 949–955.

    Article  CAS  PubMed  Google Scholar 

  26. Gasparian, M.E., Ostapchenko, V.G., Schulga, A.A., Dolgikh, D.A., and Kirpichnikov, M.P., Protein. Expr. Purif., 2003, vol. 31, no. 1, pp. 133–139.

    Article  CAS  PubMed  Google Scholar 

  27. Jenny, R.J., Mann, K.G., and Lundblad, R.L., Protein. Expr. Purif., 2003, vol. 31, no. 1, pp. 1–11.

    Article  CAS  PubMed  Google Scholar 

  28. Milton, H. and Saier, Jr., Microbe, 2006, vol. 1, no. 9, pp. 414–419.

    Google Scholar 

  29. Tan, S., Wu, W., Liu, J., Kong, Y., Pu, Y., and Yuan, R., Protein. Expr. Purif., 2002, vol. 25, no. 3, pp. 430–436.

    Article  CAS  PubMed  Google Scholar 

  30. Messens, J. and Collet, J.F., Int. J. Biochem. Cell Biol., 2006, vol. 38, no. 7, pp. 1050–1062.

    Article  CAS  PubMed  Google Scholar 

  31. Bowden, G.A. and Georgiou, G., J. Biol. Chem., 1990, vol. 265, no. 28, pp. 16760–16766.

    CAS  PubMed  Google Scholar 

  32. Rosenthal, J.A., Hsu, S.H., Schneider, D., Gentile, L.N., Messier, N.J., Vaslet, C.A., and Hawrot, E., J. Biol. Chem., 1994, vol. 269, no. 15, pp. 11178–11185.

    CAS  PubMed  Google Scholar 

  33. Li, J., Zhang, H., Liu, J., and Xu, K., Biochem. J., 2006, vol. 398, no. 2, pp. 233–242.

    Article  CAS  PubMed  Google Scholar 

  34. Gong, N., Armugam, A., and Jeyaseelan, K., Eur. J. Biochem., 1999, vol. 265, no. 3, pp. 982–989.

    Article  CAS  PubMed  Google Scholar 

  35. Drevet, P., Lemaire, C., Gasparini, S., Zinn-Justin, S., Lajeunesse, E., Ducancel, F., Pinkasfeld, S., Courcon, M., Tremeau, O., Boulain, J.C., and Menez, A., Protein. Expr. Purif, 1997, vol. 10, pp. 293–300.

    Article  CAS  PubMed  Google Scholar 

  36. Hsieh, H.C., Kumar, T.K., and Yu, C., Biochem. Biophys. Res. Commun., 2004, vol. 20, no. 4, pp. 1374–1381.

    Article  CAS  Google Scholar 

  37. Antil, S., Servent, D., and Menez, A., J. Biol. Chem., 1999, vol. 274, pp. 34851–34858.

    Article  CAS  PubMed  Google Scholar 

  38. Wang, Y., Jing, L., and Xu, K., J. Biotechnol., 2002, vol. 94, pp. 235–244.

    Article  CAS  PubMed  Google Scholar 

  39. Chang, L., Lin, J., Wu, P., Chang, C., and Hong, E., Biochem. Biophys. Res. Commun., 1997, vol. 230, no. 1, pp. 192–195.

    Article  CAS  PubMed  Google Scholar 

  40. Lyukmanova, E.N., Shulga, A.A., Arsenieva, D.A., Pluzhnikov, K.A., Dolgikh, D.A., Arseniev, A.S., and Kirpichnikov, M.R., Bioorg. Khim., 2004, vol. 30, pp. 30–40 [Russ. J. Bioorg. Chem. (Engl. Transl.), 2004, vol. 30, pp. 25—34].

    Google Scholar 

  41. Peng, L.S., Zhong, X.F., Huang, Y.S., Zhang, Y., Zheng, S.L., Wei, J.W., Wu, W.Y., and Xu, A.L., Toxicon, 2003, vol. 42, no. 7, pp. 753–761.

    Article  CAS  PubMed  Google Scholar 

  42. Schein, C.H. and Noteborn, M.H.M., Biotechnology, 1988, vol. 6, no. 3, pp. 291–295.

    Article  CAS  Google Scholar 

  43. Young, H.S., Herbette, L.G., and Skita, V., Biophys. J., 2003, vol. 85, no. 2, pp. 943–953.

    Article  CAS  PubMed  Google Scholar 

  44. Lesovoy, D.M., Bocharov, E.V., Lyukmanova, E.N., Kosinsky, Yu.A., Shulepko, M.A., Dolgikh, D.A., Kirpichnikov, M.P., Efremov, R.G., and Arseniev, A.S., Biophys. J., 2009, vol. 97, pp. 2089–2097.

    Article  CAS  PubMed  Google Scholar 

  45. Chang, L.S., Chen, K.C., Wu, B.N., Lin, S.K., Wu, P.F., Hong, Y.R., and Yang, C.C., Biochem. Biophys. Res. Commun., 1999, vol. 263, no. 3, pp. 652–656.

    Article  CAS  PubMed  Google Scholar 

  46. Lyukmanova, E.N., Shulepko, M.A., Tikhonov, R.V., Shenkarev, Z.O., Paramonov, A.S., Vulfson, A.N., Kasheverov, I.E., Ustich, T.L., Utkin, Yu.N., Arseniev, A.S., Tsetlin, V.I., Dolgikh, D.A., and Kirpichnikov, M.P., Biokhimiya, 2009, vol. 74, no. 10, pp. 1142–1149.

    CAS  Google Scholar 

  47. Fiordalisi, J.J., Rabiee, R., Chiappinelli, V.A., and Grant, G.A., Biochemistry, 1994, vol. 33, no. 13, pp. 3872–3877.

    Article  CAS  PubMed  Google Scholar 

  48. Bocharov, E.V., Lyukmanova, E.N., Ermolyuk, Ya.S., Shulga, A.A., Pluzhnikov, K.A., Dolgikh, D.A., Kirpichnikov, M.P., and Arseniev, A.S., Appl. Magn. Res., 2003, vol. 24, pp. 247–254.

    Article  CAS  Google Scholar 

  49. Krabben, L., van Rossum, B.J., Jehle, S., Bocharov, E.V., Lyukmanova, E.N., Schulga, A.A., Arseniev, A.S., Hucho, F., and Oschkinat, H., J. Mol. Biol., 2009, vol. 390, pp. 662–671.

    Article  CAS  PubMed  Google Scholar 

  50. Lyukmanova, E.N., Shenkarev, Z.O., Schulga, A.A., Ermolyuk, Y.S., Mordvintsev, D.Y., Utkin, Y.N., Shoulepko, M.A., Hogg, R.C., Bertrand, D., Dolgikh, D.A., Tsetlin, V.I., and Kirpichnikov, M.P., J. Biol. Chem., 2007, vol. 282, pp. 24784–24791.

    Article  CAS  PubMed  Google Scholar 

  51. Levandoski, M.M., Caffery, P.M., Rogowski, R.S., Lin, Y., Shi, Q.-L., and Hawrot, E., J. Neurochem., 2000, vol. 74, pp. 1279–1289.

    CAS  PubMed  Article  Google Scholar 

  52. Mourier, G., Servent, D., Zinn-Justin, S., and Menez, A., Protein. Eng., 2000, vol. 13, no. 3, pp. 217–225.

    Article  CAS  PubMed  Google Scholar 

  53. Poh, S.L., Mourier, G., Thai, R., Armugam, A., Molgo, J., Servent, D., Jeyaseelan, K., and Menez, A., Eur. J. Biochem., 2002, vol. 269, no. 17, pp. 4247–4256.

    Article  CAS  PubMed  Google Scholar 

  54. Tugarinov, V., Kanelis, V., and Kay, L.E., Nat. Protoc., 2006, vol. 1, pp. 749–754.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    E. N. Lyukmanova, M. A. Shulepko, Z. O. Shenkarev, D. A. Dolgikh & M. P. Kirpichnikov

  2. Lomonosov Moscow State University, Moscow, Russia

    M. A. Shulepko, D. A. Dolgikh & M. P. Kirpichnikov

Authors
  1. E. N. Lyukmanova
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  2. M. A. Shulepko
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  3. Z. O. Shenkarev
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  4. D. A. Dolgikh
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  5. M. P. Kirpichnikov
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Correspondence to E. N. Lyukmanova.

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Original Russian Text © E.N. Lyukmanova, M.A. Shulepko, Z.O. Shenkarev, D.A. Dolgikh, M.P. Kirpichnikov, 2010, published in Bioorganicheskaya Khimiya, 2010, Vol. 36, No. 2, pp. 149–158.

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Lyukmanova, E.N., Shulepko, M.A., Shenkarev, Z.O. et al. In vitro production of three-finger neurotoxins from snake venoms, a disulfide rich proteins. Problems and their solutions (Review). Russ J Bioorg Chem 36, 137–145 (2010). https://doi.org/10.1134/S1068162010020019

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  • DOI: https://doi.org/10.1134/S1068162010020019

Key words

  • α-neurotoxins
  • nicotinic acetylcholine receptor
  • bacterial expression
  • three-finger proteins
  • expression in Escherichia coli