Skip to main content
Log in

Two New Cysteine Endopeptidases Obtained from the Latex of Araujia hortorum Fruits

  • Published:
Journal of Protein Chemistry Aims and scope Submit manuscript

Abstract

Two new endopeptidases were purified to homogeneity from the latex of Araujia hortorum fruits by a simple purification procedure involving ultracentrifugation and ion exchange chromatography. Molecular weights of araujiain h II and araujiain h III were 23,718 and 23546 (mass spectrometry), respectively. The isoelectric point of araujiain h II was 8.9, whereas araujiain h III had a pI higher than 9.3. Maximum proteolytic activity on caseine was reached at pH 8.0-9.0 for both endopeptidases, which were irreversibly inhibited by iodoacetate and E-64, suggesting they belong to the cysteine protease family. Esterolytic activity was determined on N-α-CBZ-amino acid-p-nitrophenyl esters, and the highest k cat/K m values for the both enzymes were obtained with the glutamine derivative. The N-terminal sequences of araujiain h II and araujiain h III showed a high degree of homology with other plant cysteine endopeptidases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

REFERENCES

  • Abraham, K. I. and Joshi, P. N. (1979a). Biochim.Biophys.Acta 568, 111–119.

    Article  CAS  PubMed  Google Scholar 

  • Abraham, K. I. and Joshi, P. N. (1979b). Biochim.Biophys.Acta 568, 120–126.

    Article  CAS  PubMed  Google Scholar 

  • Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D. J. (1997). Nucleic Acids Res. 25, 3389–3402.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arribére, M. C., Cortadi, A. A., Gattuso, M. A., Bettiol, M. P., Priolo, N. S., and Caffini, N. O. (1998). Phytochem.Anal. 9, 267–273.

    Article  Google Scholar 

  • Arribére, M. C., Vairo Cavalli, S. E., Priolo, N. S., Caffini, N. O., Gattuso, M., and Cortadi, A. A. (1999). Acta Horticulturae 501, 259–268.

    Article  Google Scholar 

  • Barragán, B. E., Cruz, M. T., del Castillo, L. M., and Castañeda-Agulló, M. (1985). Rev.Latinoamer.Quím. 16, 117–119.

    Google Scholar 

  • Barrett, A. J., Rawlings, N. D., and Woessner, J. F. (1998). Handbook of Proteolytic Enzymes, Academic Press, London, pp. 3, 545–797.

    Google Scholar 

  • Boller, T. (1986). In Plant Proteolytic Enzymes (Dalling, M. J., ed.), CRC Press, Boca Raton, Florida, Vol. I, pp. 76–86.

    Google Scholar 

  • Bradford, M. M. (1976). Anal.Biochem. 72, 248–254.

    Article  CAS  PubMed  Google Scholar 

  • Brockbank, W. J. and Lynn, K. R. (1979). Biochim.Biophys.Acta 578, 113–122.

    Google Scholar 

  • Burkart, A. (1979). Flora ilustrada de Entre Ríos (Argentina), Parte V.Dicotiledóneas Metaclamídeas: Gamopétalas, Col. Cient. del INTA, Buenos Aires, pp. 173–175.

    Google Scholar 

  • Choi, K. H., Laursen, R. A., and Allen, K. N. (1999). Biochemistry 38, 11624–11633.

    Article  CAS  PubMed  Google Scholar 

  • Dixon, M. and Webb, E. C. (1979). In Enzymes, Academic Press, New York, pp. 164–169.

    Google Scholar 

  • Dubois, T., Kleinschmidt, T., Schenk, A. G., Looze, Y., and Braunitzer, G. (1988). Biol.Chem.Hoppe-Seyler 369, 741–754.

    Article  CAS  PubMed  Google Scholar 

  • Feller, U. (1986). In Plant Proteolytic Enzymes (Dalling, M. J., ed.), CRC Press, Boca Raton, Florida, Vol. II, p. 55.

    Google Scholar 

  • Good, N. E. and Izawa, S. (1972). Meth.Enzymol. 24, 53–68.

    Article  CAS  Google Scholar 

  • Jones, M. L., Larse, P. B., and Woodson, W. R. (1995). Plant Mol.Biol. 28, 505–512.

    Article  CAS  PubMed  Google Scholar 

  • Laemmli, U. K. (1970). Nature 227, 680–685.

    Article  CAS  PubMed  Google Scholar 

  • Lee, K. L., Albee, K. L., Bernasconi, R. J., and Edmunds, T. (1997). Biochem.J. 327, 199–202.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lynn, K. R., Brockbank, W. J., and Clevette-Radford, N. A. (1980). Biochim.Biophys.Acta 612, 119–125.

    Article  CAS  PubMed  Google Scholar 

  • Mitchel, R. E., Chaiken, I. M., and Smith, E. L. (1970). J.Biol.Chem. 245, 3485–3492.

    Article  CAS  PubMed  Google Scholar 

  • Pal, G. and Sinha, N. K. (1980). Arch.Biochem.Biophys. 202, 321–329.

    Article  CAS  PubMed  Google Scholar 

  • Priolo, N. S., López, L. M. I., Arribére, M. C., Natalucci, C. L., and Caffini, N. O. (1991). Acta Alimentaria 20, 189–196.

    CAS  Google Scholar 

  • Priolo, N., Morcelle del Valle, S., Arribére, M. C., López, L., and Caffini, N. (2000). J.Protein Chem. 19, 39–49.

    Article  CAS  PubMed  Google Scholar 

  • Rudenskaya, G. N., Bogacheva, A. M., Preusser, A., Kuznetsova, A. V., Dunaevsky, Ya. E., Golovkin, B. N., and Stepanov, V. M. (1998). FEBS Lett. 437, 237–240.

    Article  CAS  PubMed  Google Scholar 

  • Silverstein, R. M. (1974). Anal.Biochem. 62, 478–484.

    Article  CAS  PubMed  Google Scholar 

  • Tablero, M., Arreguín, R., Arreguín, B., Soriano, M., Sánchez, R. I., Rodríguez Romero, A., and Hernández-Arana, A. (1991). Plant Sci. 74, 7–15.

    Article  CAS  Google Scholar 

  • Taylor, M. A., Al-Sheikh, M., Revell, D. F., Sumner, I. G., and Connerton, I. F. (1999). Plant Sci. 145, 41–47.

    Article  CAS  Google Scholar 

  • Tranbarger, T. J. and Misra, S. (1996). Gene 172, 221–226.

    Article  CAS  PubMed  Google Scholar 

  • Uhlig, H. (1998). Industrial Enzymes and Their Applications, Wiley, New York, pp. 146–147.

    Google Scholar 

  • Walreavens, V., Jaziri, M., van Beeumen, J., Schnek, A. G., Kleinschmidt, T., and Looze, Y. (1993). Biol.Chem.Hoppe-Seyler 374, 501–506.

    Article  CAS  PubMed  Google Scholar 

  • Watanabe, H., Abe, K., Emori, Y., Hosoyama, H., and Arai, S. (1991). J.Biol.Chem. 266, 16897–16902.

    Article  CAS  PubMed  Google Scholar 

  • Watt, J. M. and Breyer-Brandwijk, M. G. (1962). The Medicinal and Poisonous Plants of Southern and Eastern Africa.Being an Account of Their Medicinal and Other Uses, Chemical Composition, Pharmacological Effects and Toxicology in Man and Animal, 2nd ed., Livingstone, Edinburgh.

    Google Scholar 

  • Ye, Z. H. and Varner, J. E. (1996). Plant Mol.Biol. 30, 1233–1246.

    Article  CAS  PubMed  Google Scholar 

  • Zhao, C., Johnson, B. J., Kositsup, B., and Beers, E. P. (2000). Plant Physiol. 123, 1185–1196.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Nora Priolo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Obregón, W.D., Arribére, M.C., Morcelle del Valle, S. et al. Two New Cysteine Endopeptidases Obtained from the Latex of Araujia hortorum Fruits. J Protein Chem 20, 317–325 (2001). https://doi.org/10.1023/A:1010953718679

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1010953718679

Navigation