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Glycosidase activity in the excretory-secretory products of the liver fluke, Fasciola hepatica

Published online by Cambridge University Press:  14 September 2004

J. A. IRWIN ,
P. E. W. MORRISSEY ,
J. P. RYAN ,
A. WALSHE ,
S. M. O'NEILL ,
S. D. CARRINGTON ,
E. MATTHEWS ,
E. FITZPATRICK ,
G. MULCAHY  and
A. P. CORFIELD
...Show all authors
J. A. IRWIN
Affiliation:
Department of Veterinary Physiology and Biochemistry, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
P. E. W. MORRISSEY
Affiliation:
Department of Veterinary Physiology and Biochemistry, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
J. P. RYAN
Affiliation:
Department of Veterinary Physiology and Biochemistry, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
A. WALSHE
Affiliation:
School of Biological Sciences, Dublin City University, Glasnevin, Dublin 9, Ireland
S. M. O'NEILL
Affiliation:
School of Biological Sciences, Dublin City University, Glasnevin, Dublin 9, Ireland
S. D. CARRINGTON
Affiliation:
Department of Veterinary Anatomy, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
E. MATTHEWS
Affiliation:
Department of Veterinary Anatomy, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
E. FITZPATRICK
Affiliation:
Department of Veterinary Anatomy, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
G. MULCAHY
Affiliation:
Department of Veterinary Microbiology and Parasitology, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
A. P. CORFIELD
Affiliation:
Mucin Research Group, Division of Medicine, Jenner Yard, Bristol Royal Infirmary, Marlborough Street, Bristol BS2 8HW, UK
J. P. DALTON
Affiliation:
School of Biological Sciences, Dublin City University, Glasnevin, Dublin 9, Ireland Present address: Institute for the Biotechnology of Infectious Diseases (IBID), University of Technology, Sydney, Westbourne Street, Gore Hill, Sydney, NSW 2065, Australia.
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Abstract

Fasciola hepatica secretes proteolytic enzymes and other molecules that are essential for host penetration and migration. This mixture may include enzymes required for the degradation of supramucosal gels, which defend epithelial surfaces against pathogen entry. These contain hydrated mucins that are heavily glycosylated. Excretory-secretory products (ES) from F. hepatica were examined for a range of glycosidase activities, using synthetic 4-methylumbelliferyl glycosides as substrates. The ES product contained at least 8 different glycosidase activities, the most abundant of which were β-N-acetylhexosaminidase, β-galactosidase and β-glucosidase. Alpha-fucosidase, β-glucuronidase, α-galactosidase, α-mannosidase and neuraminidase were also present. β-N-acetylhexosaminidase and β-galactosidase were present in multiple isoforms (at least 4), whereas β-glucosidase appeared to exist as one isoenzyme with a pI <3·8. All three enzymes had acidic pH optima (4·5–5·0). Ovine small intestinal mucin was degraded by ES at pH 4·5 or 7·0, with or without active cathepsin L, the major protease found in F. hepatica ES. The ability of F. hepatica ES to degrade mucin in the presence or absence of active cathepsin L suggests that cathepsin L is not essential for mucin degradation. The abundance of β-galactosidase and β-hexosaminidase in ES supports a role for these enzymes in mucin degradation.

Keywords

mucinβ-galactosidaseβ-N-acetylhexosaminidaseβ-glucosidase
Type
Research Article
Information
Parasitology , Volume 129 , Issue 4 , October 2004 , pp. 465 - 472
Copyright
2004 Cambridge University Press

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References

REFERENCES

ABDEL-RAHMAN, S., O'REILLY, K. L. & MALONE, J. B. (1999). Biochemical characterization and localization of Fasciola hepatica 26–28 kDa diagnostic coproantigen. Parasite Immunology 21, 279–286.CrossRefGoogle Scholar
BERRIN, J. G., McLAUCHLAN, W. R., NEEDS, P., WILLIAMSON, G., PUIGSERVER, A., KROON, P. A. & JUGE, N. (2002). Functional expression of human liver cytosolic beta-glucosidase in Pichia pastoris. Insights into its role in the metabolism of dietary glucosides. European Journal of Biochemistry 269, 249–258.Google Scholar
BRADFORD, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle Scholar
CARMONA, C., DOWD, A. J., SMITH, A. M. & DALTON, J. P. (1993). Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Molecular and Biochemical Parasitology 62, 9–17.CrossRefGoogle Scholar
COLLINS, P. R., STACK, C. N., O'NEILL, S. M., DOYLE, S., RYAN, T., BRENNAN, G. P., MOUSLEY, A., STEWART, M., MAULE, A. G., DALTON, J. P. & DONNELLY, S. (2004). Cathepsin L1, the major protease involved in liver fluke (Fasciola hepatica) virulence: propeptide cleavage sites and autoactivation of the zymogen secreted from gastrodermal cells. Journal of Biological Chemistry 279, 17038–17046.CrossRefGoogle Scholar
CORFIELD, A. P. & MYERSCOUGH, N. (2000). Glycosidase activity. Methods in Molecular Biology 125, 403–416.CrossRefGoogle Scholar
DALTON, J. P., McGONIGLE, S., ROLPH, T. P. & ANDREWS, S. J. (1996). Induction of protective immunity in cattle against infection with Fasciola hepatica by vaccination with cathepsin L proteinases and with hemoglobin. Infection and Immunity 64, 5066–5074.Google Scholar
DAVIES, J. R. & CARLSTEDT, I. (2000). Isolation of large gel-forming mucins. Methods in Molecular Biology 125, 3–13.Google Scholar
DOWD, A. J., DOOLEY, M., O'FÁGÁIN, C. & DALTON, J. P. (2000). Stability studies on the cathepsin L proteinase of the helminth parasite, Fasciola hepatica. Enzyme and Microbial Technology 27, 599–604.CrossRefGoogle Scholar
EVERSON PEARSE, A. G. (1972). Histochemistry, Theoretical and Applied, vol. 2, 3rd Edn, pp. 1324–1325. Churchill Livingstone, Edinburgh and London.
FREIRE, T., CASARAVILLA, C., CARMONA, C. & OSINAGA, E. (2003). Mucin-type O-glycosylation in Fasciola hepatica: characterisation of carcinoma-associated Tn and sialyl-Tn antigens and evaluation of UDP-GalNAc[ratio ]polypeptide N-acetylgalactosaminyl-transferase activity. International Journal for Parasitology 33, 47–56.CrossRefGoogle Scholar
FUJINO, T. & ISHII, Y. (1986). Comparative histochemical studies of glycosidase activity in some helminths. Journal of Helminthology 60, 1–13.CrossRefGoogle Scholar
JEFFERIES, J. R., CAMPBELL, A. M., VAN ROSSUM, A. J., BARRETT, J. & BROPHY, P. M. (2001). Proteomic analysis of Fasciola hepatica excretory-secretory products. Proteomics 1, 1128–1132.3.0.CO;2-0>CrossRefGoogle Scholar
LAEMMLI, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–685.CrossRefGoogle Scholar
LOJDA, Z. (1970). Indigogenic methods for glycosidases. II. An improved method for beta-D-galactosidase and its application to localization studies of the enzymes in the intestine and in other tissues. Histochemie 23, 266–288.Google Scholar
MAS-COMA, S., BARGUES, M. D. & ESTEBAN, J. G. (1999). Human fasciolasis. In Fasciolosis (ed. Dalton, J. P.), pp. 411–434. CAB International, Wallingford, UK.
MATERN, H., HEINEMANN, H., LEGLER, G. & MATERN, S. (1997). Purification and characterization of a microsomal bile acid beta-glucosidase from human liver. Journal of Biological Chemistry 272, 11261–11267.CrossRefGoogle Scholar
MOORE, M. N. & HALTON, D. W. (1976). Fasciola hepatica: histochemical observations on juveniles and adults and the cytopathological changes induced in infected mouse liver. Experimental Parasitology 40, 212–224.CrossRefGoogle Scholar
MULCAHY, G., O'CONNOR, F., McGONIGLE, S., DOWD, A., CLERY, D. G., ANDREWS, S. J. & DALTON, J. P. (1998). Correlation of specific antibody titre and avidity with protection in cattle immunized against Fasciola hepatica. Vaccine 16, 932–939.CrossRefGoogle Scholar
O'NEILL, S. M., MILLS, K. H. & DALTON, J. P. (2001). Fasciola hepatica cathepsin L cysteine proteinase suppresses Bordetella pertussis-specific interferon-gamma production in vivo. Parasite Immunology 23, 541–547.CrossRefGoogle Scholar
SMITH, A. M., DOWD, A. J., HEFFERNAN, M., ROBERTSON, C. D. & DALTON, J. P. (1993). Fasciola hepatica: a secreted cathepsin L-like proteinase cleaves host immunoglobulin. International Journal for Parasitology 23, 977–983.CrossRefGoogle Scholar
THORNTON, D. J., HOLMES, D. F., SHEEHAN, J. K. & CARLSTEDT, I. (1989). Quantitation of mucus glycoproteins blotted onto nitrocellulose membranes. Analytical Biochemistry 182, 160–164.CrossRefGoogle Scholar
WILKINSON, G. N. (1961). Statistical estimations in enzyme kinetics. Biochemical Journal 80, 324–332.CrossRefGoogle Scholar