Skip to main content

Specific features of enteropeptidase hydrolysis of chimeric proteins at the specific linker (Asp)4Lys depending on the refolding conditions

Abstract

Refolding from inclusion bodies of chimeric proteins containing the enteropeptidase-specific linker (Asp)4Lys was carried out. It was shown that, depending on the refolding conditions, chimeric proteins function as substrates or inhibitors of the enteropeptidase. The efficiency of the enteropeptidase hydrolysis of chimeric proteins containing the (Asp)4Lys linker may depend not only on the amino acid sequence of the protein binding site for the enzyme but also on the site’s conformation.

This is a preview of subscription content, access via your institution.

Abbreviations

HAV:

protein of the VP1 region of the hepatitis A virus

HIV:

protease of the human immunodeficiency virus

HIVR :

protease covalently joined with reverse transcriptase of the human immunodeficiency virus

Nfa:

naphthylamide

References

  1. Nygren, P.-A., Stahl, S., and Ulhen, M.,Trends Biotechnol., 1994, vol. 12, pp. 184–188.

    PubMed  Article  CAS  Google Scholar 

  2. He, M., Jin, L., and Austen, B.,J. Protein Chem., 1993, vol. 12, pp. 1–5.

    PubMed  Article  Google Scholar 

  3. Mikhailova, A.G., Shibanova, E.D., Rumsh, L.D., and Antonov, V.K.,Bioorg. Khim., 1994, vol. 20, pp. 883–893.

    PubMed  CAS  Google Scholar 

  4. Kutuzov, M.A., Shmukler, B.E., Suslov, O.N., Zargarov, A.A., and Abdulaev, N.G.,Bioorg. Khim., 1992, vol. 18, pp. 623–634.

    PubMed  CAS  Google Scholar 

  5. Gaidarov, I.O., Suslov, O.N., Ovchinnikova, T.V., and Abdulaev, N.G.,Bioorg. Khim., 1994, vol. 20, pp. 367–381.

    PubMed  CAS  Google Scholar 

  6. Kostanyan, I.A., Astapova, M.V., Starovoitova, E.V., Dranitsyna, S.M., and Lipkin, V.M.,Bioorg. Khim., 1995, vol. 21, pp. 243–248.

    CAS  Google Scholar 

  7. Hopp, T.P. and Prickett, K.S.,Bio/Technology, 1988, vol. 6, pp. 1204–1210.

    Article  CAS  Google Scholar 

  8. LaVallie, E.R. and DiBlasio, E.A.,Bio/Technology, 1993, vol. 11, pp. 187–193.

    PubMed  Article  CAS  Google Scholar 

  9. Shibanova, E.D., Rumsh, L.D., and Aleksandrov, S.L.,Tezisy IV simpoziuma “Khimiya proteoliticheskikh fermentov”, IV Simp. “Chemistry of Proteolytic Enzymes”), Moscow, 1997, p. 74.

  10. Sablina, E.P., Amerik, A.Yu., and Antonov, V.K.,Bioorg. Khim., 1991, vol. 17, pp. 1201–1212.

    PubMed  CAS  Google Scholar 

  11. Provincher, S.V.,Comput. Phys. Commun., 1982, vol. 27, pp. 229–242.

    Article  Google Scholar 

  12. Miranker, A., Robinson, C.V., Radford, S.E., and Dobson, C.M.,FASEB J., 1996, vol. 10, pp. 93–101.

    PubMed  CAS  Google Scholar 

  13. Popov, E.M.,Problema belka (The Problem of Protein), Moscow: Nauka, 1997, vol. 3, pp. 459–466.

    Google Scholar 

  14. Dill, K.A. and Chan, H.S.,Nature Struct. Biol., 1997, vol. 4, pp. 10–19.

    PubMed  Article  CAS  Google Scholar 

  15. Bessmertnaya, L.Ya., Loiko, I.I., Goncharova, T.I., Ivanov, N.V., Rumsh, L.D., and Antonov, V.K.,Biokhimiya, 1997, vol. 62, pp. 993–1000.

    Google Scholar 

  16. Light, A. and Janska, H.,Trends Biochem. Sci., 1989, vol. 14, pp. 110–112.

    PubMed  Article  CAS  Google Scholar 

  17. Mikhailova, A.G. and Rumsh, L.D.,Bioorg. Khim., 1998, vol. 24, pp. 282–287.

    PubMed  CAS  Google Scholar 

  18. Hosfield, T. and Lu, Q.,Anal. Biochem., 1999, vol. 269, pp. 10–16.

    PubMed  Article  CAS  Google Scholar 

  19. Carlson, J.D. and Yarmush, M.L.,Bio/Technology, 1992, vol. 10, pp. 86–91.

    PubMed  Article  CAS  Google Scholar 

  20. Aleksandrov, S.L.,Bioorg. Khim., 1994, vol. 20, pp. 5–13.

    PubMed  CAS  Google Scholar 

  21. Mikhailova, A.G. and Rumsh, L.D.,Appl. Biochem. Biotechnol. (in press).

  22. Schneider, C.,J. Biol. Chem., 1982, vol. 257, pp. 10766–10769.

    PubMed  CAS  Google Scholar 

  23. Eisenthal, R. and Cornish-Bowden, A.,Biochem. J., 1974, vol. 139, pp. 715–720.

    PubMed  CAS  Google Scholar 

  24. Laemmli, U.K.,Nature, 1970, vol. 227, pp. 680–685.

    PubMed  Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to E. D. Shibanova.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shibanova, E.D., Mikhailova, A.G., Aleksandrov, S.L. et al. Specific features of enteropeptidase hydrolysis of chimeric proteins at the specific linker (Asp)4Lys depending on the refolding conditions. Russ J Bioorg Chem 26, 466–473 (2000). https://doi.org/10.1007/BF02758617

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02758617

Key words

  • chimeric proteins
  • folding
  • limited proteolysis
  • enteropeptidase