Hostname: page-component-77c89778f8-gvh9x Total loading time: 0 Render date: 2024-07-19T20:39:49.735Z Has data issue: false hasContentIssue false
  • English
  • Français

Solubilization and immunoprecipitation of 125I-labelled antigens from Plasmodium knowlesi schizont-infected erythrocytes using non-ionic, anionic and zwitterionic detergents

Published online by Cambridge University Press:  06 April 2009

R. J. Howard  and
J. W. Barnwell
R. J. Howard
Affiliation:
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20205
J. W. Barnwell
Affiliation:
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20205
Get access Rights & Permissions [Opens in a new window]

Summary

Plasmodium knowlesi malaria-infected erythrocytes were radio-iodinated and several non-ionic, anionic and zwitterionic detergents were compared in their capacity to extract the labelled membrane proteins. The use of these detergents for antigen identification was tested by immunoprecipitation, after addition of Triton X-100 to some detergent extracts, using hyperimmune monkey antiserum and protein A-Sepharose. 125I-labelled antigens were specifically immunoprecipitated with all detergents tested, including the anionic detergents sodium dodecyl sulphate (SDS), deoxycholate and cholate; the zwitterions Zwittergent-312 and -314, CHAPS and Empigen BB, as well as several non-ionic detergents. The SDS-polyacrylamide gel electrophoresis patterns of 125I-labelled antigens varied after extraction with different detergents, there being no consistent pattern for detergents of a particular class. A total of 14 125I-labelled antigens were identified, 11 of them using Triton X-100. Some minor antigens identified with Triton X-100 were immunoprecipitated in greater amount after extraction in other detergents. Most importantly, two antigens Mr 200000 and 180000 were detected only after extraction with deoxycholate or SDS.

Type
Research Article
Information
Parasitology , Volume 88 , Issue 1 , February 1984 , pp. 27 - 36
Copyright
Copyright © Cambridge University Press 1984

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

Barnwell, J. W., Howard, R. J. & Miller, L. H. (1982). Altered expression of Plasmodium knowlesi variant antigen on the erythrocyte membrane is splenectomized rhesus monkeys. Journal of Immunology 128, 224–6.Google Scholar
Brown, K. N. (1974). Antigenic variation and immunity to malaria. In Parasites in the Immunized Host: Mechanisms of Survival, Ciba Fdn. Symp. 25, pp. 35–46. London: Elsevier.Google Scholar
Brown, K. N. & Brown, I. N. (1965). Immunity to malaria: Antigenic variation in chronic infections of Plasmodium knowlesi. Nature, London 208, 1286–8.Google Scholar
Deans, J. A., Dennis, E. D. & Cohen, S. (1978). Antigenic analysis of sequential erythrocytic stages of Plasmodium knowlesi. Parasitology 77, 333–44.CrossRefGoogle ScholarPubMed
Gonenne, A. & Ernst, R. (1978). Solubilization of membrane proteins by sulfobetaines, novel zwitterionic surfactants. Analytical Biochemistry 87, 28–38.CrossRefGoogle ScholarPubMed
Grant, D. A. W. & Hjerten, S. (1977). Some observations on the choice of detergent for solubilization of the human erythrocyte membrane. The Biochemical Journal 164, 465–8.CrossRefGoogle ScholarPubMed
Grant, P. T. & Fulton, J. D. (1957). The catabolism of glucose by strains of Trypanosoma rhodesiense. The Biochemical Journal 66, 242–50.CrossRefGoogle ScholarPubMed
Hjelmeland, L. M. (1980). A nondenaturing zwitterionic detergent for membrane biochemistry: Design and synthesis. Proceedings of the National Academy of Sciences, USA 77, 6368–70.Google Scholar
Howard, R. J., Barnwell, J. W., Kao, V., Daniel, W. A. & Aley, S. B. (1982). Radioiodination of new protein antigens on the surface of Plasmodium knowlesi schizont-infected erythrocytes. Molecular and Biochemical Parasitology 6, 343–67.Google Scholar
Howard, R. J. & Kao, V. (1982). Comparison of the surface membrane proteins of human and rhesus monkey (Macaca mulatta) erythrocytes labelled with proteins and glycoprotein radiolabelling probes. Comparative Biochemistry and Physiology 70B, 767–74.Google Scholar
Johnson, J. G., Epstein, N., Shiroishi, T. & Miller, L. H. (1981). Identification of surface proteins on viable Plasmodium knowlesi merozoites. Journal of Protozoology 28, 160–9.CrossRefGoogle ScholarPubMed
Laemmli, U. K. & Favre, M. (1973). Maturation of the head of bacteriophage T4. I. DNA packaging events. Journal of Molecular Biology 80, 575–99.Google Scholar
Liljas, L., Lundahl, P. & Hjerten, S. (1974). Selective solubilization with Tween 20 of proteins from water-extracted human erythrocyte membranes. Analysis by gel electrophoresis in dodecylsulfate and in Tween 20. Biochimica et biophysica acta 352, 327–37.CrossRefGoogle ScholarPubMed
Scheibel, L. W. & Miller, J. (1969). Cytochrome oxidase activity in platelet-free preparations of Plasmodium knowlesi. Journal of Parasitology 55, 825–9.CrossRefGoogle ScholarPubMed
Schmidt-Ulrich, R. & Wallach, D. F. H. (1978). Plasmodium knowlesi-induced antigens in membranes of parasitized rhesus monkey erythrocytes. Proceedings of the National Academy of Sciences, USA 75, 4949–53.CrossRefGoogle Scholar
Swanstrom, R. & Shank, P. R. (1978). X-ray intensifying screens greatly enhance the detection by autoradiography of the radioactive isotypes 32P and 125I. Analytical Biochemistry 86, 184–92.Google Scholar
Yu, J., Fischman, D. A. & Steck, T. L. (1973). Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. Journal of Supramolecular Structure 1, 233–48.CrossRefGoogle ScholarPubMed