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Isolation and partial characterization of the tegumental outer membrane of adult Schistosoma mansoni

Published online by Cambridge University Press:  06 April 2009

A. J. G. Simpson ,
M. D. Schryer ,
I. M. Cesari ,
W. H. Evans  and
S. R. Smithers
A. J. G. Simpson
Affiliation:
Divisions of Parasitology, National institute for Medical Research, Mill Hill, London NW7 1AA
M. D. Schryer
Affiliation:
Divisions of Parasitology, National institute for Medical Research, Mill Hill, London NW7 1AA
I. M. Cesari
Affiliation:
Divisions of Parasitology, National institute for Medical Research, Mill Hill, London NW7 1AA
W. H. Evans
Affiliation:
Divisions of BiochemistryNational institute for Medical Research, Mill Hill, London NW7 1AA
S. R. Smithers
Affiliation:
Divisions of Parasitology, National institute for Medical Research, Mill Hill, London NW7 1AA
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Summary

Incubation of adult Schistosoma mansoni at 37 °C in chemically defined media caused the shedding of variable amounts of the parasite's tegument. In phosphate-buffered saline, pH 7·4, 37% of 125I-labelled wheat germ agglutinin which was attached to the schistosome surface was released within 5 min. When the released material was collected by centrifugation at 55000 g for 1 h, the pellet was seen to consist of components of the parasite tegument and contained 1% of the parasite protein and 6·2% of the 5′-AMPase activity. By monitoring the release of the gut enzyme, haemoglobinase, it was shown that predominantly tegumental material was released under these conditions. The material released from the parasite was separated by discontinuous sucrose gradient centrifugation into 4 major fractions and it was shown that a material banding at low density was highly enriched in vesicles of parasite outer membrane. Major polypeptides of apparent molecular weights 135000, 110000, 90000, 65000, 47000 and 40000 and glycoproteins of apparent molecular weights 135000, 120000, 65000, 63000 and 30–40000 were identified in this fraction. Comparison of the distribution of 5′-AMPase activity and the contents of each fraction as revealed by ultrastructural examination showed that this enzyme is a good marker for the tegumental outer membrane.

Type
Research Article
Information
Parasitology , Volume 83 , Issue 1 , August 1981 , pp. 163 - 177
Copyright
Copyright © Cambridge University Press 1981

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References

REFERENCES

Avruch, J. & Wallach, D. F. H. (1971). Preparation and properties of plasma membrane and endoplasmic reticulum fragments from isolated rat fat cells. Biochimica et biophyica acta 233, 334–47.CrossRefGoogle ScholarPubMed
Bogitsch, B. J. & Krupa, P. L. (1971). Schistosoma mansoni and Haematoloechus medioplexus: Nucleosidediphosphatase localisation in the tegument. Experimental Parasitology 30, 418–25.CrossRefGoogle Scholar
Chang, K-J., Bennett, V. & Cuatrecasas, P. (1975). Membrane receptors as general markers for plasma membrane isolation procedures. Journal of Biological Chemistry 250, 488–500.CrossRefGoogle ScholarPubMed
Cordeiro, M. N. & Gazzinelli, G. (1979). Schistosoma mansoni: resolution and molecular weight estimates of tegument glycoproteins by polyacrylamide gel electrophoresis. Experimental Parasitology 48, 337–44.CrossRefGoogle ScholarPubMed
Cotman, C. W. & Matthews, D. A. (1971). Synaptic plasma membranes from rat brain synaptosomes. Isolation and partial characterization. Biochimica et biophysica acta 249, 380–94.CrossRefGoogle ScholarPubMed
Ernst, S. C. (1977). Biochemical and cytochemical studies of alkaline phosphatase activity in Schistosoma mansoni. Rice University Studies 62, 81–95.Google Scholar
Evans, W. H. (1978). Preparation and characterisation of mammalian plasma membranes. In Laboratory Techniques in Biochemistry and Molecular Biology, vol. 7, Part 1 (ed. Work, T. S. and Work, E.), pp. 1–266. North Holland.Google Scholar
Evans, W. H. & Bruning, J. W. (1970). Studies on the distribution of some histocompatibility antigens in mouse liver plasma membrane and microsomal fractions. Immunology 19, 735–41.Google ScholarPubMed
Gurd, J. W. & Evans, W. H. (1976). Identification of plasma membrane glycoproteins which bind to 125I labelled Con A following electrophoresis in SDS. Canadian Journal of Biochemistry 54, 477–80.CrossRefGoogle Scholar
Hayunga, E. G., Murrell, K. D., Taylor, D. W. & Vannier, W. E. (1979 a). Isolation and characterisation of surface antigens from Schistosoma mansoni I. Evaluation of techniques for radioisotope labelling of surface proteins from adult worms. Journal of Parasitology 65, 488–96.CrossRefGoogle ScholarPubMed
Hayunga, G. H., Murrell, K. D., Taylor, D. W. & Vannier, W. E. (1979 b). Isolation and characterisation of surface antigens from Schistosoma mansoni II. Antigenicity of radio-labelled proteins from adult worms. Journal of Parasitology 65, 497–506.CrossRefGoogle Scholar
Hirsch, J. G. & Fedorko, M. E. (1968). Ultrastructure of human leukocytes after simultaneous fixation with gluteraldehyde and osmium-tetroxide and ‘post-fixation’ in uranyl acetate. Journal of Cell Biology 38, 615–27.CrossRefGoogle Scholar
Hockley, D. J. & McLaren, D. J. (1973). Schistosoma mansoni: changes in the outer membrane of the tegument during development from cercariae to adult worm. International Journal for Parasitology 3, 13–25.CrossRefGoogle ScholarPubMed
Hunter, W. M. & Greenwood, F. C. (1962). Preparation of iodine-131 labelled human growth of high specific activity. Nature, London 194, 495–6.CrossRefGoogle ScholarPubMed
Kusel, J. R. (1972). Protein composition and protein synthesis in the surface membranes of Schistosoma mansoni. Parasitology 65, 55–9.CrossRefGoogle ScholarPubMed
Kusel, J. R., Sher, A., Perez, H., Clegg, J. A. & Smithers, S. R. (1975). Use of radioactive isotopes in the study of specific schistosome membrane antigens. In Nuclear Techniques in Helminthology Research, pp. 127–43. International Atomic Energy Agency, Vienna.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lowry, O. H., Rosebrough, M. S., Fare, A. L. & Randall, R. J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 193, 265–75.CrossRefGoogle ScholarPubMed
McLaren, D. J. (1974). The anterior glands of adult Necator americanus (Nematoda: Strongyloidea). I. Ultrastructural studies. International Journal for Parasitology 4, 25–37.CrossRefGoogle ScholarPubMed
Morris, G. P. & Threadgold, L. T. (1968). Ultrastructure of the tegument of adult Schistosoma mansoni. Journal of Parasitology 54, 15–27.CrossRefGoogle ScholarPubMed
Murrell, K. D., Taylor, D. W., Vannier, W. E. & Dean, D. E. (1978). Schistosoma mansoni: analysis of surface membrane carbohydrates using lectins. Experimental Parasitology 46, 247–55.CrossRefGoogle ScholarPubMed
Nicolson, G. L. (1976). Transmembrane control of the receptors on normal and tumour cells. 1. Cytoplasmic influence over cell surface components. Biochimica et biophysica acta 457, 57–108.CrossRefGoogle Scholar
Ramasamy, R. (1979). Surface proteins on schistosomula and cercariae of Schistosoma mansoni. International Journal for Parasitology 9, 491–3.CrossRefGoogle ScholarPubMed
Rosen, H. (1957). A modified ninhydrin colorimetric analysis for amino acids. Archives of Biochemistry and Biophysics 67, 10–15.CrossRefGoogle ScholarPubMed
Ruppel, A. (1978). A study of surface proteins in the parasite Schistosoma mansoma. Ph.D. thesis, University of Freiburg, GFR.Google Scholar
Simpson, A. J. G. & Smithers, S. R. (1980). Characterization of the exposed carbohydrates on the surface membrane of adult Schistosoma mansoni by analysis of lectin binding. Parasitology 81, 1–15.CrossRefGoogle ScholarPubMed
Smith, J. H., Reynolds, E. S. & Lichtenberg, F. Von (1969). The integument of Schistosoma mansoni. American Journal of Tropical Medicine and Hygiene 18, 28–49.CrossRefGoogle ScholarPubMed
Smithers, S. R. & Terry, R. J. (1965). The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of adult worms. Parasitology 55, 695–700.CrossRefGoogle ScholarPubMed
Studier, F. W. (1973). Analysis of bacteriophage T7 early RNAs and proteins on slab gels. Journal of Molecular Biology 79, 237–48.CrossRefGoogle ScholarPubMed
Timms, A. R. & Bueding, E. (1959). Studies of a proteolytic enzyme from Schistosoma mansoni. British Journal of Pharmacology 14, 68–73.Google ScholarPubMed
Tulkens, P., Beaufay, H. & Trouet, A. (1974). Analytical fractionation of homogenates from cultured rat embryo fibroblasts. Journal of Cell Biology 63, 383–401.CrossRefGoogle ScholarPubMed
Wisher, M. H. & Evans, W. H. (1975). Functional polarity of the rat hepatocyte surface membrane. The Biochemical Journal 146, 375–88.CrossRefGoogle ScholarPubMed