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Activities of elements of the yeast α-factor precursor leader at different stages of somatropin secretion by Saccharomyces cerevisiae

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Abstract

An analytical system for a quantitative assessment of contributions of pre- and pro-components of the yeast α-factor leader to key stages of somatropin secretion in Saccharomyces cerevisiae has been designed. Using this system, it was shown that: 1—the α-factor pro-peptide has contributed almost equally to the secretion at the Erv29p-dependent stage of vesicular transport and at the translocation stage; 2—of the two above stages, the Erv29p-dependent stage could only be optimized by using of two copies of the α-factor pro-peptide; 3—when the α-factor pre-peptide and the artificial pro-peptide with significantly greater hydrophobicity were compared it was found that somatropin secretion apparently depended on the translocation mode as higher secretion was achieved with hydrophobic artificial pre-peptide associated with the co-translational translocation.

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Abbreviations

ER:

endoplasmic reticulum

PAGE:

polyacrylamide gel electrophoresis

References

  1. Idiris, A., Tohda, H., Kumagai, H., and Takegawa, K., Engineering of protein secretion in yeast: strategies and impact on protein production, Appl. Microbiol. Biotechnol., 2010, vol. 86, pp. 403–417.

    Article  CAS  PubMed  Google Scholar 

  2. Platt, K., Mothes, W., Wilkinson, B.M., Stirling, C.J., and Rapoport, T.A., Signal sequence recognition in posttranslational protein transport across the yeast ER membrane, Cell, 1998, vol. 94, pp. 795–807.

    Article  Google Scholar 

  3. Chaudhuri, B., Steube, K., and Stephan, C., The proregion of the yeast pre-pro-α-factor is essential for membrane translocation of human insulin-like growth factor1 in vitro, Eur. J. Biochem., 1992, vol. 206, pp. 793–800.

    Article  CAS  PubMed  Google Scholar 

  4. Becker, J., Walter, W., Yan, W., and Craig, E.A., Functional interaction of cytosolic hsp70 and a DnaJ-related protein, Ydjlp, in protein translocation in vivo, Mol. Cell. Biol., 1996, vol. 16, pp. 4378–4386.

    CAS  PubMed Central  PubMed  Google Scholar 

  5. Belden, W.J. and Barlowe, C., Role of Erv29p in collecting soluble secretory proteins into ER-derived transport vesicles, Science, 2001, vol. 264, pp. 1528–1531.

    Article  Google Scholar 

  6. Otte, S. and Barlowe, C., Sorting signals can direct receptor-mediated export of soluble proteins into COPII vesicles, Nat. Cell Biol., 2004, vol. 6, pp. 1189–1194.

    Article  CAS  PubMed  Google Scholar 

  7. Tritos, N.A. and Mantzoros, C.S., Recombinant human growth hormone: old and novel uses, Am. J. Med., 1998, vol. 105, pp. 44–57.

    Article  CAS  PubMed  Google Scholar 

  8. Rosenfeld, R.G., Cohen, P., Robison, L.L., Bercu, B.B., Clayton, P., Hoffman, A.R., Radovick, S., Saenger, P., Savage, M.O., and Wit, J.M., Long-term surveillance of the growth hormone therapy, J. Clin. Endocrinol. Metab., 2012, vol. 97, pp. 68–72.

    Article  CAS  PubMed  Google Scholar 

  9. Kartasheva, N.N., Kuchin, S.V., and Benevolensky, S.V., Genetic aspects of carbon catabolite repression of the STA2 glucoamylase gene in Saccharomyces cerevisiae, Yeast, 1996, vol. 12, pp. 1297–1300.

    Article  CAS  PubMed  Google Scholar 

  10. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Lab. Press, 1989.

    Google Scholar 

  11. Kazachenko, K.J., Kozlov, D.G., Ezdakov, I.V., Gubajdullin, I.I., Glazunov, A.V., Cheperegin, S.E., and Efremov, B.D., Method for microbiological synthesis of secreted human somatotropin and yeast strain Saccharomyces cerevisiae-secreted human somatotropin producer, RF Patent No. RU2460795, 2012

    Google Scholar 

  12. Ito, Y., Fukuda, Y., Murata, K., and Kimura, A., Transformation of intact yeast cells treated with alkali cations J. Bacteriol., 1983, vol. 153, pp. 163–168.

    CAS  PubMed Central  PubMed  Google Scholar 

  13. Piper, P.W. and Curran, P.G., When a glycolytic gene on a yeast 2ori-stb plasmid is made essential for growth its expression level is a major determinant of plasmid copy number, Curr. Genet., 1990, vol. 17, pp. 119–123.

    Article  CAS  PubMed  Google Scholar 

  14. Tijssen, T., Practice and Theory of Enzyme Immunoassays, Amsterdam, New York, Oxford: Elsevier, 1985.

    Google Scholar 

  15. Kohara, A., Yamamoto, Y., and Kikuchi, M., Processing and secretion of human growth hormone with artificial signal sequence, Biosci. Biotechnol. Biochem., 1994, vol. 58, pp. 779–781.

    Article  CAS  PubMed  Google Scholar 

  16. Kyte, J. and Doolittle, R.F., A simple method for displaying the hydropathic character of a protein, J. Mol. Biol., 1982, vol. 157, pp. 105–132.

    Article  CAS  PubMed  Google Scholar 

  17. Ng, D.T.W., Braun, J.D., and Walter, P., Signal sequences specify the targeting route to the endoplasmic reticulum membrane, J. Cell Biol., 1996, vol. 134, pp. 269–278.

    Article  CAS  PubMed  Google Scholar 

  18. Johnsson, N. and Varshavsky, A., Ubiquitin-assisted dissection of protein transport across membranes, EMBO J., 1994, vol. 13, pp. 2686–2698.

    CAS  PubMed Central  PubMed  Google Scholar 

  19. Kozlov, D.G. and Yagudin, T.A., Antibody fragments may by incorrectly processed in the yeast Pichia pastoris, Biotehcnol. Lett., 2008, vol. 30, pp. 1661–1663.

    Article  CAS  Google Scholar 

  20. Degryse, E., Dietrich, M., Nguyen, M., Achstetter, T., Charlier, M., Charpigny, G., Gaye, P., and Martal, J., Addition of a dipeptide spacer significantly improves secretion of ovine trophoblast interferon in yeast, Gene, 1992, vol. 118, pp. 47–53.

    Article  CAS  PubMed  Google Scholar 

  21. Tsiomenko, A.B., Tuymetova, G.P., Eldarov, M.A., Korolev, S.V., Skryabin, K.G., and Kulaev, I.S., The prosegment of yeast α-factor directs a heterologous protein (human growth hormone) to the culture medium of Saccharomyces cerevisiae, Biochemistry (Moscow), 1994, vol. 59, pp. 1247–1256.

    Google Scholar 

  22. Arnold, C.E., Parekh, R.N., Yang, W., and Wittrup, K.D., Leader peptide efficiency correlates with signal recognition particle dependence in Saccharomyces cerevisiae, Biotechnol. Bioeng., 1998, vol. 59, pp. 286–293.

    Article  CAS  PubMed  Google Scholar 

  23. Swanton, E. and Bulleid, N.J., Protein folding and translocation across the endoplasmic reticulum membrane (review), Mol. Membr. Biol., 2003, vol. 20, pp. 99–104.

    Article  CAS  PubMed  Google Scholar 

  24. Rapoport, T.A., Protein translocation across the eukaryotic endoplasmic reticulum and bacterial plasma membranes, Nature, 2007, vol. 450, pp. 663–669.

    Article  CAS  PubMed  Google Scholar 

  25. Wiren, K.M., Potts, J.T., Jr., and Kronenberg, H.M., Importance of the propeptide sequence of human preproparathyroid hormone for signal sequence function, J. Biol. Chem., 1988, vol. 263, pp. 19771–19777.

    CAS  PubMed  Google Scholar 

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Correspondence to D. G. Kozlov.

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Kazachenko, K.Y., Efremov, B.D. & Kozlov, D.G. Activities of elements of the yeast α-factor precursor leader at different stages of somatropin secretion by Saccharomyces cerevisiae . Appl Biochem Microbiol 50, 829–834 (2014). https://doi.org/10.1134/S000368381409004X

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  • DOI: https://doi.org/10.1134/S000368381409004X

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