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Evidence for concanavalin A binding sites on the surface coat of Trypanosoma congolense

Published online by Cambridge University Press:  06 April 2009

P. Rautenberg
Affiliation:
Institut für Veterinär-Biochemie, Freie Universität Berlin, Koserstrasse 20, 1000 Berlin 33, W. Germany
E. Reinwald
Affiliation:
Institut für Veterinär-Biochemie, Freie Universität Berlin, Koserstrasse 20, 1000 Berlin 33, W. Germany
H-J. Risse
Affiliation:
Institut für Veterinär-Biochemie, Freie Universität Berlin, Koserstrasse 20, 1000 Berlin 33, W. Germany

Summary

Glycoproteins of Trypanosoma congolense have been detected on SDS-polyacrylamide gels using the Concanavalin A peroxidase technique. Using [35S]diazoniobenzenesulphonate as a marker for cell surface proteins it was possible to distinguish between internal glycoproteins and the surface coat proteins. On SDS-polyacrylamide gels Con A reacted with the surface coat proteins. Results obtained from Con A-induced agglutination of living trypanosomes indicated that sugars of the surface coat proteins were accessible to Con A. This was reinforced by the cytochemical visualization of Con A binding to the trypanosome surface. The results suggested that the surface coat protein contained α-linked d-mannosyl, d-glucosyl, or N-acetyl-d-glucosaminoyl residues, which are exposed exteriorly on the surface coat.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1980

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References

Baltz, T., Baltz, D. & Pautrizel, R. (1976). Affinité de la concanavaline A pour Trypanosoma equiperdum: applications à l'isolement de la fraction glycoprotéique spécifique du type antigénique. Annales d'Immunologie (Inst. Pasteur) 127 C, 761–74.Google Scholar
Baltz, T., Baltz, D., Pautrizel, R., Lambin, G. & Degand, P. (1977). Chemical and immunological characterisation of specific glycoproteins from Trypanosoma equiperdum variants. Febs Letters 82, 93–6.CrossRefGoogle ScholarPubMed
Berg, H. C. & Hirsh, D. (1975). Synthesis of diazotized 35S-sulfanilic acid of high specific activity: a label for the outer surface of cell membranes. Analytical Biochemistry 66, 629–31.CrossRefGoogle ScholarPubMed
Bernhard, W. & Avrameas, S. (1971). Ultrastructural visualization of cellular carbohydrate components by means of concanavalin A. Experimental Cell Research 64, 232–6.CrossRefGoogle ScholarPubMed
Bonner, W. M. & Laskey, R. A. (1974). A film detection method for tritium labelled proteins and nucleic acids in polyacrylarnide gels. European Journal of Biochemistry 46, 83–8.CrossRefGoogle ScholarPubMed
Bridgen, P. J., Cross, G. A. M. & Bridgen, J. (1976). N-terminal amino acid sequences of variant-specific surface antigens from Trypanosoma brucei. Nature, London 263, 613–14.CrossRefGoogle ScholarPubMed
Cross, G. A. M. (1975). Identification, purification and properties of clone-specific glycoprotein antigen constituting the surface coat of Trypanosoma brucei. Parasitology 71, 393417.CrossRefGoogle ScholarPubMed
Cross, G. A. M. & Johnson, J. G. (1976). Structure and organisation of the variant-specific antigens of Trypanosoma brucei. In Biochemistry of Parasites and Host–Parasite Relationships (ed. van den Bossche, H.), pp. 413420. Amsterdam, New York and Oxford: North-Holland Publishing Company.Google Scholar
Diffley, P., Honigberg, B. M. & Mohn, F. A. (1976). An improved method of cryopreservation of Trypanosoma (Nannomonas) congolense Broden in liquid nitrogen. Journal of Parasitology 62, 136–7.CrossRefGoogle ScholarPubMed
Doyle, J. J. (1977). Antigenic variation in the salivarian trypanosomes. In Advances in Experimental Medicine and Biology, vol. 93 (ed. Miller, L. H., Pino, J. A. and McKelvey, J. J.), pp. 2763. London and New York: Plenum Press.Google Scholar
Fairbanks, G., Steck, T. L. & Wallach, D. F. H. (1971). Electrophoretic analysis of the major polypeptides of the erythrocyte membrane. Biochemistry 10, 2606–17.CrossRefGoogle ScholarPubMed
Goldstein, I. J. (1975). Studies on the combining sites of concanavalin A. In Advances in Experimental Medicine and Biology, vol. 55, (ed. Chowdhury, T. K. and Weiss, A. K.), pp. 3553. New York and London: Plenum Press.Google Scholar
Goldstein, I. J. (1976). Carbohydrate binding specificity of concanavalin A. In Concanavalin A as a Tool, (ed. Bittinger, H. and Schnebli, H. P.), pp. 5565. London: Wiley.Google Scholar
Gray, A. R. & Luckins, A. G. (1976). Antigenic variation in salivarian trypanosomes. In Biology of the Kinetoplastida, vol. 1, (ed. Lumsden, W. H. R. and Evans, D. A.), pp. 493542. London and New York: Academic Press.Google Scholar
Johnson, J. G. & Cross, G. A. M. (1977). Carbohydrate composition of variant-specific surface antigen glycoproteins from Trypanosoma brucei. Journal of Protozoology 24, 587–91.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lanham, S. M. & Godfrey, D. G. (1970). Isolation of salivarian trypanosomes from man and other mammals using DEAE-cellulose. Experimental Parasitology 28, 521–34.CrossRefGoogle ScholarPubMed
Lis, H. & Sharon, N. (1973). The biochemistry of plant lectins (phytohemagglutinins). Annual Review of Biochemistry 42, 541–74.CrossRefGoogle ScholarPubMed
McKenzie, G. H. & Sawyer, W. H. (1973). The binding properties of dimeric and tetrameric concanavalin A. Journal of Biological Chemistry 248, 549–56.CrossRefGoogle ScholarPubMed
McKenzie, G. H., Sawyer, W. H. & Nichol, L. W. (1972) The molecular weight and stability of concanavalin A. Biochimica et Biophysica Acta 263, 283–93.CrossRefGoogle ScholarPubMed
Raadt, P. de (1974). Immunity and antigenic variation: clinical observations suggestive of immune phenomena in African trypanosomiasis. In Ciba Fdn Symp. 20 N.S., pp. 199224. Amsterdam: Associate Scientific Publishers.Google Scholar
Reinwald, E., Risse, H-J. & Sälker, R. (1978). Diazoniobenzenesulfonate as a marker for cell surface proteins: study of the surface coat of Trypanosoma congolense. Hoppe-Seyler's Zeitschrift für physiologische Chemie 359, 939–44.CrossRefGoogle ScholarPubMed
Reinwald, E., Rautenberg, P. & Risse, H-J. (1978). Characterization and isolation of the surface antigen of Trypanosoma congolense. Hoppe Seyler's Zeitschrift für physiologische Chemie 359, 1138.Google Scholar
Reinwald, E., Rautenberg, P. & Risse, H-J. (1979). Trypanosoma congolense: mechanical removal of the surface coat in vitro. Experimental Parasitology (in the Press).CrossRefGoogle ScholarPubMed
Rovis, L., Barbet, A. F. & Williams, R. O. (1978). Characterization of the surface coat of Trypanosoma congolense. Nature, London 271, 654–6.CrossRefGoogle ScholarPubMed
Seed, T. M., Seed, J. R. & Brindley, D. (1976). Surface properties of bloodstream trypanosomes (Trypanosoma brucei brucei). Tropenmedizin und Parasitologie 27, 202–12.Google Scholar
Steiger, R. F. (1975). Ultracytochemistry of the surface coat/pellicle complex in Trypanosoma brucei. Acta Tropica 32, 152–8.Google ScholarPubMed
Vickerman, K. (1974). Antigenic variation in African trypanosomes. In Ciba Fdn Symp. 25, N.S., pp. 5380. Amsterdam: Associated Scientific Publishers.Google Scholar
Vickerman, K. (1978). Antigenic variation in trypanosomes. Nature, London 273, 613–17.CrossRefGoogle ScholarPubMed
Wood, J. G. & Sarinana, F. O. (1975). The staining of sciatic nerve glycoproteins on polyacrylamide gels with concanavalin A–peroxidase. Analytical Biochemistry 69, 320–2.CrossRefGoogle ScholarPubMed
Wright, K. A. & Hales, H. (1970). Cytochemistry of the pellicle of bloodstream forms of Trypanosoma (Trypanozoon) brucei. Journal of Parasitology 56, 671–83.CrossRefGoogle ScholarPubMed