Hostname: page-component-848d4c4894-sjtt6 Total loading time: 0 Render date: 2024-06-30T17:44:24.953Z Has data issue: false hasContentIssue false
  • English
  • Français

Identification of isolate-specific proteins on sorbitol-enriched Plasmodium falciparum infected erythrocytes from Gambian patients

Published online by Cambridge University Press:  06 April 2009

S. B. Aley ,
J. A. Sherwood ,
K. Marsh ,
O. Eldelman  and
R. J. Howard
S. B. Aley
Affiliation:
The Malaria Section, Laboratory of Parasitic Diseases, National Institute of Health, Bethesda, MD 20205, USA
J. A. Sherwood
Affiliation:
The Malaria Section, Laboratory of Parasitic Diseases, National Institute of Health, Bethesda, MD 20205, USA
K. Marsh
Affiliation:
Medical Research Council Laboratories Fajara, The Gambia
O. Eldelman
Affiliation:
Division of Cancer Biology and Diagnosis, National Cancer Institute, National Institutes of Health, Bethesda, MD 20205, USA
R. J. Howard
Affiliation:
The Malaria Section, Laboratory of Parasitic Diseases, National Institute of Health, Bethesda, MD 20205, USA
Get access Rights & Permissions [Opens in a new window]

Summary

We have compared the surface radio-iodinated proteins of uninfected and Plasmodium falciparum-infected erythrocytes from natural infections of human patients. Cryopreserved infected blood from Gambian children with falciparum malaria was thawed, cultured to the middle trophozoite stage, and surface radio-iodinated. Trophozoite-infected cells were enriched about 10-fold on a Percoll gradient newly designed to separate cells based on their differential permeability to sorbitol. Infected blood was radio-iodinated and erythrocyes from the fraction enriched in parasitized cells and uninfected erythrocytes from the same sample obtained from the gradient and compared by SDS–PAGE and autoradiography. In each sample, parasitized erythrocytes contained one or more polypeptides of very high molecular weight (Mr 250000–300000) that were not found on uninfected erythrocytes from the same patient. These proteins were isolate-specific in size and number, suggesting that natural isolates contain a variable number of different P. falciparum phenotypes for this surface protein. In addition, these radio-iodinated surface proteins could not be extracted from the host cell membrane by the non-ionic detergent Triton X-100, but were extracted by SDS. The properties of these proteins suggest they are the equivalent for natural infections of the strain-dependent antigen previously described (Leech, Barnwell, Miller & Howard, 1984) on the surface of P. falciparum-infected Aotus erythrocytes. In addition, we observed a second parasite-dependent modification of labelled proteins on infected erythrocytes with the appearance of a new band of Mr 30000. There were also variations in the pattern of radio-isotope labelled proteins on uninfected erythrocytes from different patients.

Type
Research Article
Information
Parasitology , Volume 92 , Issue 3 , June 1986 , pp. 511 - 525
Copyright
Copyright © Cambridge University Press 1986

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

Aley, S. B., Barnwell, J. W., Daniel, W. A. & Howard, R. J. (1984). Identification of parasite proteins in a membrane preparation enriched for the surface membrane of erythrocytes infected with Plasmodium knowlesi. Molecular and Biochemical Parasitology 12, 69–84.CrossRefGoogle Scholar
Aley, S. B., Sherwood, J. A. & Howard, R. J. (1984). Knob-positive and knob-negative Plasmodium falciparum differ in expression of a strain-specific malarial antigen on the surface of infected erythrocytes. Journal of Experimental Medicine 160, 1585–90.CrossRefGoogle ScholarPubMed
Burkot, T. R., Williams, J. L. & Schneider, I. (1984). Infectivity to mosquitoes of Plasmodium falciparum clones grown in vitro from the same isolate. Transactions of the Royal Society of Tropical Medicine and Hygiene 78, 339–41.CrossRefGoogle ScholarPubMed
Ginsburg, H., Krugliak, M., Eidelman, O. & Cabantchik, Z. I. (1983). New permeability pathways induced in membranes of Plasmodium falciparum-infected erythrocytes. Molecular and Biochemical Parasitology 8, 177–90.CrossRefGoogle ScholarPubMed
Green, T. J., Gadsen, A., Seed, T., Jacobs, R., Morhardt, M. & Brackett, R. (1985). Cloning and characterization of Plasmodium falciparum FCR-/FMG strain. American Journal of Tropical Medicine and Hygiene 34, 24–30.CrossRefGoogle ScholarPubMed
Howard, R. J. & Barnwell, J. W. (1985). Immunochemical analysis of surface membrane antigens on erythrocytes infected with non-cloned SICA [+] or cloned SICA [-] Plasmodium knowlesi. Parasitology 92, 245–61.CrossRefGoogle Scholar
Howard, R. J., Barnwell, J. W. & Kao, V. (1983). Antigenic variation in Plasmodium knowlesi malaria: Identification of the variant antigen on infected erythrocytes. Proceedings of the National Academy of Sciences, USA 80, 4129–33.CrossRefGoogle ScholarPubMed
Howard, R. J., Barnwell, J. W., Kao, V., Daniel, W. A. & Aley, S. B. (1982). Radio-iodination of a new protein antigen on the surface of Plasmodium knowlesi schizont-infected erythrocytes. Molecular and Biochemical Parasitology 6, 343–67.CrossRefGoogle Scholar
Kramer, K. J., Chow Kan, S. & Siddiqui, W. A. (1982). Concentration of Plasmodium falciparum infected erythrocytes by density gradient centrifugation in Percoll. Journal of Parasitology 68, 336–7.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
Lambros, C. & Vanderberg, J. P. (1979). Synchronization of Plasmodium falciparum erythrocytic stages in culture. Journal of Parasitology 65, 418–20.CrossRefGoogle ScholarPubMed
Leech, J. H., Barnwell, J. W., Miller, L. H. & Howard, R. J. (1984). Identification of a strain-specific malarial antigen exposed on the surface of Plasmodium falciparum-infected erythrocytes. Journal of Experimental Medicine 159, 1567–75.CrossRefGoogle ScholarPubMed
Meryman, H. T. & Hornblower, M. (1972). A method for freezing and washing red blood cells using a high glycerol concentration. Transfusion 12, 145–56.CrossRefGoogle ScholarPubMed
Mrema, J. E., Campbell, R., Miranda, R., Jaranillo, A. L. & Rieckmann, K. H. (1979). Concentration and separation of erythrocytes infected with Plasmodium falciparum by gradient centrifugation. Bulletin of the World Health Organization 54, 133–8.Google Scholar
Pavia, C. S., Diggs, C. L. & Williams, J. (1983). The use of metrizamide for isopycnic separation and enrichment of Plasmodium falciparum schizonts from continuous culture. American Journal of Tropical Medicine and Hygiene 32, 675–81.CrossRefGoogle ScholarPubMed
Rivadeneira, E. M., Wasserman, M. & Epinal, C. T. (1983). Separation and concentration of schizonts of Plasmodium falciparum by Percoll gradients. Journal of Protozoology 30, 367–70.CrossRefGoogle ScholarPubMed
Saul, A., Myler, P. E. & Kidson, C. T. (1982). Purification of mature schizonts of Plasmodium falciparum on colloidal silica gradients. Bulletin of the World Health Organization 60, 755–9.Google ScholarPubMed
Siddiqui, W. A., Schnell, J. V. & Richmond-Crum, S. (1974). In vitro cultivation of Plasmodium falciparum at high parasitemia. American Journal of Tropical Medicine and Hygiene 23, 1015–18.CrossRefGoogle ScholarPubMed
Trager, W. & Jensen, J. B. (1976). Human malaria parasites in continuous culture. Science 193, 673–5.CrossRefGoogle ScholarPubMed
Udeinya, I. J., Graves, P. M., Carter, R., Aikawa, M. & Miller, L. H. (1983). Plasmodium falciparum: Effect of time in continuous culture on binding to human endothelial cells and amelanotic melanoma cells. Experimental Parasitology 56, 207–14.CrossRefGoogle ScholarPubMed
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