Hostname: page-component-78c5997874-94fs2 Total loading time: 0 Render date: 2024-11-03T00:50:45.326Z Has data issue: false hasContentIssue false

Characterization of proteinases in extracts of adult Haemonchus contortus, the ovine abomasal nematode

Published online by Cambridge University Press:  06 April 2009

D. P. Knox
Affiliation:
Moredun Research Institute, 408 Gilmerton Road, Edinburgh EH17 7JH
D. L. Redmond
Affiliation:
Moredun Research Institute, 408 Gilmerton Road, Edinburgh EH17 7JH
D. G. Jones
Affiliation:
Moredun Research Institute, 408 Gilmerton Road, Edinburgh EH17 7JH

Summary

The degradation of several protein substrates, including the blood proteins haemoglobin, albumin and fibrinogen, by proteinases present in extracts of adult Haemonchus contortus was examined over a broad pH range. These proteinases were further characterized on the basis of substrate specificity, inhibitor sensitivity and molecular size by spectrophotometric and substrate gel analysis. The majority of the proteinases capable of degrading the blood proteins tested were active at acidic pH and could be ascribed to the cysteine proteinase class. In addition, evidence is presented that these proteinases are differentially recognized and inhibited by immune sera and that parasites capable of withstanding protective host immune responses exhibit modified expression of proteinases.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1993

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Barrett, A. J. (1977). Introduction to the history and classification of tissue proteinases. In Proteinases in Mammalian Cells and Tissues (ed, Barrett, A. J.), pp. 118. Amsterdam, New York, Oxford: North Holland Publishing Company.Google Scholar
Bogitsch, B. J. & Dresden, M. H. (1983). Fluorescent biochemistry of acid proteases in adult Schistosoma mansoni and Schistosoma japonicum. Journal of Parasitology 69, 106–10.CrossRefGoogle Scholar
Bogitsch, B. J. & Kirschner, K. F. (1986). Schistosoma japonicum: ultrastructural localisation of a pepstatin sensitive haemoglobinase using mercury labelled pepstatin. Experimental Parasitology 62, 211–15.Google Scholar
Britton, C., Knox, D. P., Urquhart, G. & Kennedy, M. w. (1992). Characterisation of proteinases of Dictyocaulus viviparus, and their inhibition by antibody from infected and vaccinated calves. Parasitology 105, 325–33.CrossRefGoogle Scholar
Chappell, c. L. & Dresden, M. H. (1986). Schistosoma mansoni: proteinase activity of ‘haemoglobinase’ from the digestive tract of adult worms. Experimental Parasitology 61, 160–7.CrossRefGoogle Scholar
Cox, G. N., Milhausen, M. & Hageman, R. (1990 a). Anticoagulant and anthelmintic proteins and methods of production for the same. European Patent Application No. 0 434 909 A2.Google Scholar
Cox, G. N., Pratt, D., Hageman, R. & Boisvenue, R. J. (1990 b). Molecular cloning and primary sequence of a cysteine proteinase expressed by Haemonchus contortus adult worms. Molecular and Biochemical Parasitology 41, 2534.CrossRefGoogle ScholarPubMed
Hotez, P. J. & Cerami, A. (1983). Secretion of a proteolytic anticoagulant by Ancylostoma hookworms. Journal of Experimental Medicine 157, 15941603.CrossRefGoogle ScholarPubMed
Hotez, P. J., Letrang, N., McKerrow, J. H. & Cerami, A. (1985). Isolation and characterisation of a proteolytic enzyme from the adult hookworm Ancylostoma caninum. Journal of Biological Chemistry 260, 7343–8.CrossRefGoogle ScholarPubMed
Jasmer, D. P. & McGuire, T. C. (1991). Protective immunity to a blood-feeding nematode (Haemonchus contortus) induced by parasite gut antigens. Infection and Immunity 59, 4412–17.CrossRefGoogle ScholarPubMed
Jones, v. E. & Ogilvie, B. M. (1972). Protective immunity to Nippostrongylus brasiliensis in the rat. III. Modulation of worm acetylcholinesterase by antibodies. Immunology 22, 119–29.Google Scholar
Knox, D. P. & Jones, D. G. (1990). Studies on the presence and release of proteolytic enzymes (proteinases) in gastro-intestinal nematodes of ruminants. International Journal for Parasitology 20, 243–9.CrossRefGoogle ScholarPubMed
Knox, D. P. & Jones, D. G. (1992). A comparison of superoxide dismutase (EC 1.15.1.1) distribution in gastro-intestinal nematodes. International Journal for Parasitology 22, 209–14.Google Scholar
Knox, D. P. & Kennedy, M. w. (1988). Proteinases released by the parasitic larval stages of Ascaris suum and their inhibition by antibody. Molecular and Biochemical Parasitology 28, 207–16.CrossRefGoogle ScholarPubMed
McKerrow, J. H. & Doenhoff, M. J. (1988). Schistosome proteases. Parasitology Today 4, 334–40.Google Scholar
Matthews, B. E. (1977). The passage of larval helminths through tissue barriers. In Parasite Invasion, Symposia of the British Society for Parasitology, vol. 15, (ed.Taylor, A. E. R. & Muller, R.), pp. 93119. Oxford: Blackwell Scientific Publications.Google Scholar
Pratt, D., Cox, G. N., Milhausen, M. J. & Boisvenue, R. J.(1990). A developmentally regulated cysteine protease gene family in Haemonchus contortus. Molecular and Biochemical Parasitology 43, 181–92.CrossRefGoogle ScholarPubMed
Pratt, D., Armes, L. G., Hageman, R., Reynolds, V., Boisvenue, R. J. & Cox, G. N. (1992). Cloning and sequence comparisons of four distinct cysteine proteases expressed by Haemonchus contortus adult worms. Molecular and Biochemical Parasitology 51, 209–18.CrossRefGoogle ScholarPubMed
Pritchard, D. I., McKean, P. G., Tighe, P. J. & Quinell, R. J. (1992). Immunology, biochemistry and molecular biology of hookworm antigens. In Parasitic Helminths: Antigens, Membranes and Genes (ed. Kennedy, M. W.), pp. 140169. London: Taylor and Francis Ltd.Google Scholar
Sakanari, J. A., Staunton, C. E., Eakin, A. E., Craik, C. S. & McKerrow, J. H. (1989). Serine proteases from nematode and protozoan parasites: Isolation of sequence homologs using generic molecular probes. Proceedings of the National Academy of Sciences, USA 86, 4863–7.Google Scholar
Senft, A. W. (1976). Observations on the physiology of the gut of Schistosoma mansoni. In Biochemistry of Parasites and Host–Parasite Relationships (ed. van den Bossche, H.) pp. 4357. Amsterdam: Elsevier/North Holland.Google Scholar
Smith, T. S. & Munn, E. A. (1990). Strategies for vaccination against gastro-intestinal nematodes. Review Scientific and Technique Office International Epizoology 9, 577–95.Google Scholar
Smith, W. D. (1992). Protection in lambs immunised with Haemonchus contortus gut membrane proteins. Research in Veterinary Science (in the Press).Google Scholar
Suttle, N. F., Knox, D. P., Angus, K. W., Jackson, F. & Coop, R. L. (1992). Effects of dietary molybdenum on nematode and host during Haemonchus contortus infection in lambs. Research in Veterinary Science 52, 230–5.CrossRefGoogle ScholarPubMed