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Purification, Properties, and N-Terminal Amino Acid Sequence of a Kallikrein-like Enzyme from the Venom of Lachesis muta rhombeata (Bushmaster)

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Abstract

Pit viper venoms contain multiple proteinases which cause considerable damage in tissues and systemic effects after envenomation. A proteinase, kallikrein-like enzyme, belonging to the serine group must play a very important role on systemic effects. The corresponding enzyme from Lachesis muta rhombeata venom was purified to homogeneity by a combination of isoelectrofocusing fractionation followed by one step of gel filtration HPLC. The enzyme focused with pI 5.0–6.5, it had a molecular mass of 32 kDa by gel filtration HPLC, had edematogenic activity, and induced a hypotensic effect in anesthetized rats. It exhibited strong N-α-tosyl-L-Arg methyl esterase (955.38 units/mg) and N-BZ-DL-Arg-pNA amidolytic (233.02 units/mg) activities, hydrolyzed tripeptide nitroanilide derivatives weakly or not at all, and cleaved selectively the A-α and B-β chains of fibrinogen, apparently leaving the Y-chain unaffected. The 30 N-terminal amino acid sequence of the L. m. rhombeata protein showed greatest identity (74% in 26 amino acids) with Crotalus viridis kallikrein-like protein, but significant similarities in sequence were observed with enzymes from other snake venoms and pig pancreatic kallikrein.

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REFERENCES

  • Aguiar, A. S., Alves, C. R., Melgarejo, A. R., and Giovanni De Simone, S. (1996) Toxicon 34, 555–565.

    Article  CAS  PubMed  Google Scholar 

  • Bjarnason, J. B., Barish, A., Direnzo, G. S., Campbell, R., and Fox, J. W. (1983). J. Biol. Chem. 258, 12566–12573.

    Article  CAS  PubMed  Google Scholar 

  • Da Silva, N. J., Aird, S. D., Seebart, C., and Kaiser, I. I. (1989). Toxicon 27, 763–771.

    Article  CAS  PubMed  Google Scholar 

  • Dinz, M. R., and Oliveira, E. B. (1992). Toxicon 30, 247–258.

    Article  Google Scholar 

  • Erlanger, B. F., Kokowsky, N., and Cohen, W. (1961). Arch. Biochem. Biophys. 95, 271–278.

    Article  CAS  PubMed  Google Scholar 

  • Ferreira, S. H. (1979). J. Pharmacol. 3, 648–658.

    Google Scholar 

  • Giovanni De Simone S., Santos, R., Araujo, M. F., and Pinho, R. T. (1994). J. Chromatogr. A 688, 356–362.

    Google Scholar 

  • Haad, J. S. (1980). Mem. Inst. Butantan 44/45, 403–423.

    Google Scholar 

  • Itoh, N., Tamaka, N., Mihashi, S., and Yamashina, I. (1987). J. Biol. Chem. 262, 3132–3135.

    Article  CAS  PubMed  Google Scholar 

  • Komori, Y., Nakai, T., and Sugihara, H. (1988). Biochim. Biophys. Acta 967, 92–102.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Lomonte, B. (1985). Toxicon 23, 173–176.

    Article  CAS  PubMed  Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). J. Biol. Chem. 193, 265–275.

    Article  CAS  PubMed  Google Scholar 

  • Markland, F. S., Kettner, C., Schiffman, S., Shaw, E., Bajwa, S. S., Readdy, K. N. N., Kirakossian, H., Paktos, G. B., Theodor, I., and Pirkle, H. (1982). Proc. Natl. Acad. Sci. USA 79, 1688–1692.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Miranda, T. L. S., Ramos, C. H. I., Freire, R. T. S., Souza, E. P., Rogana, E., Santoro, M. M., and Figueiredo, A. F. S. (1995). Braz. J. Med. Biol. Res. 28, 505–512.

    CAS  PubMed  Google Scholar 

  • Ouchterlony, O., and Nilson, L. A. (1973). In Handbook of Experimental Immunology, 2nd ed., (Weiss, D. M., ed.), Blackwell Scientific, Oxford, Chapter 19.

    Google Scholar 

  • Pirkle, H., Markland, F. S., Theodor, I., Baumgartener, R., Bajwa, S. S., and Kirakossian, H. (1981). Biochem. Biophys. Res. Commun. 99, 715–721.

    Article  CAS  PubMed  Google Scholar 

  • Queiroz, L. S., Santos Neto, H., Assukura, M. T., Reichl, A. P., and Mandelbaum, F. R. (1985). Toxicon 23, 341–345.

    Article  CAS  PubMed  Google Scholar 

  • Sanches, E. F., Magalhães, A., and Diniz, C. R. (1987). Toxicon 25, 611–619.

    Article  Google Scholar 

  • Schwartz, M. W., and Bieber, A. L. (1985). Toxicon 23, 255–269.

    Article  CAS  PubMed  Google Scholar 

  • Selistre, H. S., Queiroz, L. S., Cunha, O. A. B., De Souza, G. E. P., and Giglio, J. R. (1990). Toxicon 28, 261–273.

    Article  CAS  PubMed  Google Scholar 

  • Shieh, T. C., Kawabata, S., Kihara, H., Ohno, M., and Iwanaga, S. (1988). J. Biochem. 103, 596–605.

    Article  CAS  PubMed  Google Scholar 

  • Silva, L. M., Dihiz, C. R., and Magalhães, A. (1985). Toxicon 23, 707–718.

    Article  CAS  PubMed  Google Scholar 

  • Silva-Haad, J. (1982). Mem. Inst. Butantan 44/45, 403–423.

    Google Scholar 

  • Tschesche, H., Mair, G., Godec, G., Fiedler, F., Ehret, W., Hirschaver, C., Lemon, M., and Fritz, H. (1979). In Kinins II: Biochemistry Pathophysiology and Clinical Aspects (Fujii, S., Moriya, H., and Suzuki, T., eds.), Plenum Press, New York, Part A, p. 245.

    Chapter  Google Scholar 

  • Worthy, K., Figueroa, C. D., Dieppe, P. A., and Bhoola, K. D. (1990). Int. J. Exp. Pathol. 71, 587–601.

    CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Laboratório de Microseqüênciamento de Proteinas, Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz-FIOCRUZ, 21040-900, Rio de Janeiro, RJ, Brazil

    Salvatore Giovanni-De-Simone & Katia Novellino

  2. Departamento de Biologia Celular e Molecular, Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil

    Salvatore Giovanni-De-Simone

  3. Instituto Vital Brazil, Niteról, RJ, Brazil

    Aniesse S. Aguiar, Anibal R. Gimenez & Roberto S. de Moura

  4. Departamento de Farmacologia, Universidade Estadual do Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Roberto S. de Moura

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  1. Salvatore Giovanni-De-Simone
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  2. Aniesse S. Aguiar
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  3. Anibal R. Gimenez
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  4. Katia Novellino
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  5. Roberto S. de Moura
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Correspondence to Salvatore Giovanni-De-Simone.

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Giovanni-De-Simone, S., Aguiar, A.S., Gimenez, A.R. et al. Purification, Properties, and N-Terminal Amino Acid Sequence of a Kallikrein-like Enzyme from the Venom of Lachesis muta rhombeata (Bushmaster). J Protein Chem 16, 809–818 (1997). https://doi.org/10.1023/A:1026372018547

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  • DOI: https://doi.org/10.1023/A:1026372018547

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