Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Feb;80(4):1087–1091. doi: 10.1073/pnas.80.4.1087

Isolation and characterization of the plasma membrane of human erythrocytes infected with the malarial parasite Plasmodium falciparum.

J Gruenberg, I W Sherman
PMCID: PMC393533  PMID: 6341989

Abstract

Human erythrocytes infected with the malarial parasite Plasmodium falciparum were labeled metabolically with a mixture of 15 radioactive amino acids. When synchronously growing parasites were at the schizont stage of development infected cells were concentrated and purified by using a Percoll-Hypaque gradient. The plasma membrane of the infected erythrocyte, isolated by binding cells to a solid support (Affi-Gel 731, Bio-Rad), was less than 1% contaminated with parasite membranes. Erythrocyte membrane proteins were analyzed by polyacrylamide gel electrophoresis and autoradiography. Despite the high sensitivity of the procedure, there was no evidence for the insertion of parasite proteins into the infected host cell membrane. One possible exception is a Mr 230,000 parasite protein present maximally as 9,000 copies per infected erythrocyte membrane. Moreover, no differences in the membrane proteins were observed between a highly knobby clone and a knobless clone of the same strain of P. falciparum. These findings appear to rule out the presence of parasite protein(s) playing a structural role in the formation of knobs on the erythrocyte surface and question whether the antigenic determinants on the P. falciparum-infected erythrocyte are of parasite origin or whether such antigens represent newly exposed or chemically modified erythrocyte determinants.

Full text

PDF
1087

Images in this article

1088Image
on p.1088
1088Image
on p.1088
1089Image
on p.1089
1089Image
on p.1089
1090Image
on p.1090

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Branton D., Cohen C. M., Tyler J. Interaction of cytoskeletal proteins on the human erythrocyte membrane. Cell. 1981 Apr;24(1):24–32. doi: 10.1016/0092-8674(81)90497-9. [DOI] [PubMed] [Google Scholar]
  3. Chaimanee P., Yuthavong Y. Phosphorylation of membrane proteins from Plasmodium berghei-infected red cells. Biochem Biophys Res Commun. 1979 Apr 13;87(3):953–959. doi: 10.1016/0006-291x(79)92049-7. [DOI] [PubMed] [Google Scholar]
  4. Cohen C. M., Kalish D. I., Jacobson B. S., Branton D. Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells. J Cell Biol. 1977 Oct;75(1):119–134. doi: 10.1083/jcb.75.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gahmberg C. G., Andersson L. C. Identification and characterization of normal and malignant human blood leukocytes by surface glycoprotein patterns. Ann N Y Acad Sci. 1978 Jun 20;312:240–255. doi: 10.1111/j.1749-6632.1978.tb16806.x. [DOI] [PubMed] [Google Scholar]
  6. Jacobson B. S., Branton D. Plasma membrane: rapid isolation and exposure of the cytoplasmic surface by use of positively charged beads. Science. 1977 Jan 21;195(4275):302–304. doi: 10.1126/science.831278. [DOI] [PubMed] [Google Scholar]
  7. Jacobson B. S. Imporved method for isolation of plasma membrane on cationic beads. Membranes from Dictyostelium discoideum. Biochim Biophys Acta. 1980 Aug 14;600(3):769–780. doi: 10.1016/0005-2736(80)90479-4. [DOI] [PubMed] [Google Scholar]
  8. Kilejian A., Abati A., Trager W. Plasmodium falciparum and Plasmodium coatneyi: immunogenicity of "knob-like protrusions" on infected erythrocyte membranes. Exp Parasitol. 1977 Jun;42(1):157–164. doi: 10.1016/0014-4894(77)90073-x. [DOI] [PubMed] [Google Scholar]
  9. Kilejian A. Characterization of a protein correlated with the production of knob-like protrusions on membranes of erythrocytes infected with Plasmodium falciparum. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4650–4653. doi: 10.1073/pnas.76.9.4650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Königk E., Mirtsch S. Plasmodium chabaudi-infection of mice: specific activities of erythrocyte membrane-associated enzymes and patterns of proteins and glycoproteins of erythrocyte membrane preparations. Tropenmed Parasitol. 1977 Mar;28(1):17–22. [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lambros C., Vanderberg J. P. Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol. 1979 Jun;65(3):418–420. [PubMed] [Google Scholar]
  13. Langreth S. G., Reese R. T. Antigenicity of the infected-erythrocyte and merozoite surfaces in Falciparum malaria. J Exp Med. 1979 Nov 1;150(5):1241–1254. doi: 10.1084/jem.150.5.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schmidt-Ullrich R., Wallach D. F., Lightholder J. Metabolic labelling of P. knowlesi-specific glycoproteins in membranes of parasitized rhesus monkey erythrocytes. Cell Biol Int Rep. 1980 Jun;4(6):555–561. doi: 10.1016/0309-1651(80)90021-1. [DOI] [PubMed] [Google Scholar]
  15. Schmidt-Ullrich R., Wallach D. F., Lightholder J. Two Plasmodium knowlesi-specific antigens on the surface of schizont-infected Rhesus monkey erythrocytes induce antibody production in immune hosts. J Exp Med. 1979 Jul 1;150(1):86–99. doi: 10.1084/jem.150.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schmidt-Ullrich R., Wallach D. F. Plasmodium knowlesi-induced antigens in membranes of parasitized rhesus monkey erythrocytes. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4949–4953. doi: 10.1073/pnas.75.10.4949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sherman I. W. Biochemistry of Plasmodium (malarial parasites). Microbiol Rev. 1979 Dec;43(4):453–495. doi: 10.1128/mr.43.4.453-495.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Trager W., Jensen J. B. Human malaria parasites in continuous culture. Science. 1976 Aug 20;193(4254):673–675. doi: 10.1126/science.781840. [DOI] [PubMed] [Google Scholar]
  19. Trager W., Rudzinska M. A., Bradbury P. C. The fine structure of Plasmodium falciparum and its host erythrocytes in natural malarial infections in man. Bull World Health Organ. 1966;35(6):883–885. [PMC free article] [PubMed] [Google Scholar]
  20. Trigg P. I., Hirst S. I., Shakespeare P. G., Tappenden L. Labelling of membrane glycoprotein in erythrocytes infected with Plasmodium knowlesi. Bull World Health Organ. 1977;55(2-3):205–209. [PMC free article] [PubMed] [Google Scholar]
  21. Vettore L., De Matteis M. C., Zampini P. A new density gradient system for the separation of human red blood cells. Am J Hematol. 1980;8(3):291–297. doi: 10.1002/ajh.2830080307. [DOI] [PubMed] [Google Scholar]
  22. Weidekamm E., Wallach D. F., Lin P. S., Hendricks J. Erythrocyte membrane alterations due to infection with Plasmodium berghei. Biochim Biophys Acta. 1973 Nov 16;323(4):539–546. doi: 10.1016/0005-2736(73)90162-4. [DOI] [PubMed] [Google Scholar]
  23. Yuthavong Y., Wilairat P., Panijpan B., Potiwan C., Beale G. H. Alterations in membrane proteins of mouse erythrocytes infected with different species and strains of malaria parasites. Comp Biochem Physiol B. 1979;63(1):83–85. doi: 10.1016/0305-0491(79)90238-4. [DOI] [PubMed] [Google Scholar]
  24. van der Meulen J. A., Emerson D. M., Grinstein S. Isolation of chromaffin cell plasma membranes on polycationic beads. Biochim Biophys Acta. 1981 May 20;643(3):601–615. doi: 10.1016/0005-2736(81)90356-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES