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Characterization of three neutral proteases of Spirometra mansoni plerocercoid

Published online by Cambridge University Press:  06 April 2009

Y. Kong ,
Y.-B. Chung ,
S.-Y. Cho ,
S.-H. Choi  and
S.-Y. Kang
Y. Kong
Affiliation:
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea
Y.-B. Chung
Affiliation:
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea
S.-Y. Cho
Affiliation:
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea
S.-H. Choi
Affiliation:
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea
S.-Y. Kang
Affiliation:
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea
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Summary

In the pathogenesis of sparganosis, proteases have been considered to play important roles in tissue migration and parasite feeding. Several bands of proteolysis were observed when crude extracts of Spirometra mansoni plerocercoid (sparganum) were examined using gelatin substrate gel at neutral pH, of which two proteases of 198 and 104 kDa were purified by two chromatographic steps, and a 36 kDa protease was purified by gelatin-affinity and DEAE–anion exchange chromatography. All the purified proteases exhibited optimal activity at pH 7·5 and 0·1 M Tris–HCI. Proteolytic activities at 198 and 104 kDa were inhibited specifically by serine protease inhibitors, and 4-(amidinophenyl)methansulfonyl fluoride (APMSF, 0·5 m) and N-α–p–tosyl-L-lysine chloromethyl ketone (TLCK, 1 mM), which strongly suggested that these two proteases were trypsin-like proteases. The activity of the 36 kDa protease was inhibited by N-tosyl-l-phenylalanine chloromethyl ketone (TPCK, I mM) and chymostatin (0·1 mM), and was potentiated in 10 mM Ca2+ which showed that the protease had a chymotrypsin-like property. All the proteases were Schiff(PAS) positive. Proteases of 198 and 104 kDa degraded collagen completely within 24 h. The 36 kDa enzyme cleaved human recombinant interferon-γ (rIFNγ) and bovine myelin basic protein. In addition, all the purified proteins elicited strong antibody responses in the infected patients.

Keywords

Spirometra mansoni plerocercoidsparganosistrypsin-like proteasechymotrypsin-like protease
Type
Research Article
Information
Parasitology , Volume 108 , Issue 3 , April 1994 , pp. 359 - 368
Copyright
Copyright © Cambridge University Press 1994

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References

REFERENCES

Alexander, C. M. & Werb, Z. (1989). Proteinases and extracellular matrix remodeling. Current Opinion in Cell Biology 1, 974–82.CrossRefGoogle ScholarPubMed
Barrett, A. J. (1986). Serine proteinases. In Proteinase Inhibitors, 1st edn. (ed. Barrett, A. J. & Salvesen, G.), pp. 181–208. Amsterdam: Elsevier Science Publishers.Google Scholar
Bond, J. S. & Butler, P. E. (1987). Intracellular proteases. Annual Review of Biochemistry 56, 333–64.Google Scholar
Chang, K. H., Chi, J. C., Cho, S. Y., Han, M. H., Han, D. H. & Han, M. C. (1992). Cerebral sparganosis: Analysis of 34 cases with emphasis on CT features. Neuroradiology 34, 1–8.Google Scholar
Chang, K. H., Cho, S. Y., Chi, J. G., Kim, W. S., Han, M. C., Myung, H. & Choi, K. S. (1987). Cerebral sparganosis: CT characteristics. Radiology 165, 505–10.Google Scholar
Chappell, C. L. & Dresden, M. H. (1987). Purification of cysteine proteinase from adult Schistosoma mansoni. Archives of Biochemistry and Biophysics 256, 560–8.Google Scholar
Chi, J. G., Chi, H. S. & Lee, S. H. (1980). Histopathologic study on human sparganosis. Korean Journal of Parasitology 18, 15–23.CrossRefGoogle Scholar
Cho, S. Y. (1987). Diphyllobothriasis and sparganosis. In Oxford Textbook of Medicine, 2nd edn, (ed. Weatherall, D. J., Ledingham, J. G. G. & Warrell, D. A.), pp. 5·569–5·571. Oxford: Oxford University Press.Google Scholar
Cho, S. Y., Chung, Y. B. & Kong, Y. (1992). Component proteins and protease activities in excretory–secretory product of sparganum. Korean Journal of Parasitology 30, 227–30.Google Scholar
Cho, S. Y., Kang, S. Y. & Kong, Y. (1990). Purification of antigenic protein of sparganum by immunoaffinity chromatography using a monoclonal antibody. Korean Journal of Parasitology 28, 135–42.Google Scholar
Choi, S. H., Kang, S. Y., Kong, Y. & Cho, S. Y. (1988). Antigenic protein fractions reacting with sera of sparganosis patients. Korean Journal of Parasitology 26, 163–7.CrossRefGoogle Scholar
Dresden, M. H., Rege, A. A. & Murell, K. D. (1985). Strongyloides ransomi: proteolytic enzymes from larvae. Experimental Parasitology 59, 257–63.Google Scholar
Etges, R., Bouvier, J. & Bordier, C. (1986). The major surface protein of Leishmania promastigotes is a protease. Journal of Biological Chemistry 261, 9098–101.CrossRefGoogle ScholarPubMed
Fukase, T., Matsuda, Y., Akihama, S. & Itagaki, H. (1985). Purification and some properties of cysteine protease of Spirometra erinacei plerocercoid (Cestoda: Diphyllobothriidae). Japanese Journal of Parasitology 34, 351–60.Google Scholar
Glossmann, H. & Neville, D. M. Jr (1971). Glycoproteins of cell surface: a comparative study of three different cell surfaces of the rat. Journal of Biological Chemistry 246, 6339–46.CrossRefGoogle Scholar
Healer, J., Ashall, F. & Maizels, R. (1991). Characterization of proteolytic enzymes from larval and adult Nippostrongylus brasiliensis. Parasitology 103, 305–14.CrossRefGoogle ScholarPubMed
Kim, L. S., Kong, Y., Kang, S. Y. & Cho, S. Y. (1992). Immunohistochemical localization of 36 and 29 kDa proteins in sparganum. Korean Journal of Parasitology 30, 25–31.Google Scholar
Kong, Y., Kang, S. Y. & Cho, S. Y. (1991). Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography. Korean Journal of Parasitology 29, 1–7.Google Scholar
Kwa, B. H. (1972). Studies on the sparganum of Spirometra erinacei. II. Proteolytic enzymes in the scolex. International Journal for Parasitology 2, 29–33.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–75.Google Scholar
Maki, J., Furuhashi, A. & Yanagisawa, T. (1982). The activity of acid proteases hydrolysing haemoglobin parasitic helminths with special reference to interspecific and intraspecific distribution. Parasitology 84, 137–47.Google Scholar
McKerrow, J. H. (1989). Parasite proteinases. Experimental Parasitology 68, 111–15.Google Scholar
McKerrow, J. H. & Doenhoff, M. J. (1988). Schistosome proteases. Parasitology Today 4, 334–40.Google Scholar
McKerrow, J. H., Piano-Heiss, S., Lindquist, R. & Werb, Z. (1985). Purification and characterization of elastinolytic proteinases secreted by cercariae of Schistosoma mansoni. Journal of Biological Chemistry 260, 3703–7.Google Scholar
Müller, J. F. (1938). Studies on Sparganum mansonoides and Sparganum proliferum. American Journal of Tropical Medicine 18, 303–28.Google Scholar
Phares, C. K. (1987). Plerocercoid growth factor: a homologue of human growth hormone. Parasitology Today 3, 346–9.CrossRefGoogle ScholarPubMed
Sakanari, J. A., Staunton, C. E., Eakin, A. E., Craik, C. S. & McKerrow, J. H. (1989). Serine proteases from nematode and protozoan parasite: Isolation of sequence homologies using genetic molecular probes. Proceedings of the National Academy of Sciences, USA 86, 4863–7.CrossRefGoogle ScholarPubMed
Sauer, M. C. & Senft, A. W. (1972). Properties of a proteolytic enzyme from Schistosoma mansoni. Comparative Biochemistry and Physiology 42B, 205–20.Google Scholar
Song, C. Y., Choi, D. H., Kim, T. S. & Lee, S. H. (1992). Isolation and partial characterization of cysteine proteinase from sparganum. Korean Journal of Parasitology 30, 191–200.CrossRefGoogle ScholarPubMed
Tsang, V. C. W., Peralta, J. M. & Simons, A. R. (1983). Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods in Enzymology 92, 377–91.CrossRefGoogle ScholarPubMed