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. 1986 Oct 1;103(4):1269–1277. doi: 10.1083/jcb.103.4.1269

Secretion of a malarial histidine-rich protein (Pf HRP II) from Plasmodium falciparum-infected erythrocytes

PMCID: PMC2114335  PMID: 3533951

Abstract

Plasmodium falciparum-infected erythrocytes (IRBCs) synthesize several histidine-rich proteins (HRPs) that accumulate high levels of [3H]histidine but very low levels of amino acids such as [3H]isoleucine or [35S]methionine. We prepared a monoclonal antibody which reacts specifically with one of these HRPs (Pf HRP II) and studied the location and synthesis of this protein during the parasite's intracellular growth. With the knob-positive Malayan Camp strain of P. falciparum, the monoclonal antibody identified a multiplet of protein bands with major species at Mr 72,000 and 69,000. Pf HRP II synthesis began with immature parasites (rings) and continued through the trophozoite stage. The Mr 72,000 band of Pf HRP II, but not the faster moving bands of the multiplet, was recovered as a water-soluble protein from the culture supernatant of intact IRBCs. Approximately 50% of the total [3H]histidine radioactivity incorporated into the Mr 72,000 band was extracellular between 2 and 24 h of culture. Immunofluorescence and cryothin-section immunoelectron microscopy localized Pf HRP II to several cell compartments including the parasite cytoplasm, as concentrated "packets" in the host erythrocyte cytoplasm and at the IRBC membrane. Our results provide evidence for an intracellular route of transport for a secreted malarial protein from the parasite through several membranes and the host cell cytoplasm.

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Selected References

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  1. Aley S. B., Sherwood J. A., Howard R. J. Knob-positive and knob-negative Plasmodium falciparum differ in expression of a strain-specific malarial antigen on the surface of infected erythrocytes. J Exp Med. 1984 Nov 1;160(5):1585–1590. doi: 10.1084/jem.160.5.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  4. Hadley T. J., Leech J. H., Green T. J., Daniel W. A., Wahlgren M., Miller L. H., Howard R. J. A comparison of knobby (K+) and knobless (K-) parasites from two strains of Plasmodium falciparum. Mol Biochem Parasitol. 1983 Nov;9(3):271–278. doi: 10.1016/0166-6851(83)90102-0. [DOI] [PubMed] [Google Scholar]
  5. Howard R. J., Barnwell J. W. Solubilization and immunoprecipitation of 125I-labelled antigens from Plasmodium knowlesi schizont-infected erythrocytes using non-ionic, anionic and zwitterionic detergents. Parasitology. 1984 Feb;88(Pt 1):27–36. doi: 10.1017/s0031182000054317. [DOI] [PubMed] [Google Scholar]
  6. Howard R. J., Kao V., Barnwell J. W. Protein antigens of Plasmodium knowlesi clones of different variant antigen phenotype. Parasitology. 1984 Apr;88(Pt 2):221–237. [PubMed] [Google Scholar]
  7. Kilejian A. A unique histidine-rich polypeptide from the malaria parasite, Plasmodium lophurae. J Biol Chem. 1974 Jul 25;249(14):4650–4655. [PubMed] [Google Scholar]
  8. 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]
  9. Kilejian A. The biosynthesis of the knob protein and a 65 000 dalton histidine-rich polypeptide of Plasmodium falciparum. Mol Biochem Parasitol. 1984 Jun;12(2):185–194. doi: 10.1016/0166-6851(84)90134-8. [DOI] [PubMed] [Google Scholar]
  10. Köhler G., Howe S. C., Milstein C. Fusion between immunoglobulin-secreting and nonsecreting myeloma cell lines. Eur J Immunol. 1976 Apr;6(4):292–295. doi: 10.1002/eji.1830060411. [DOI] [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. Leech J. H., Barnwell J. W., Aikawa M., Miller L. H., Howard R. J. Plasmodium falciparum malaria: association of knobs on the surface of infected erythrocytes with a histidine-rich protein and the erythrocyte skeleton. J Cell Biol. 1984 Apr;98(4):1256–1264. doi: 10.1083/jcb.98.4.1256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leech J. H., Barnwell J. W., Miller L. H., Howard R. J. Identification of a strain-specific malarial antigen exposed on the surface of Plasmodium falciparum-infected erythrocytes. J Exp Med. 1984 Jun 1;159(6):1567–1575. doi: 10.1084/jem.159.6.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Meryman H. T., Hornblower M. A method for freezing and washing red blood cells using a high glycerol concentration. Transfusion. 1972 May-Jun;12(3):145–156. doi: 10.1111/j.1537-2995.1972.tb00001.x. [DOI] [PubMed] [Google Scholar]
  15. Ravetch J. V., Feder R., Pavlovec A., Blobel G. Primary structure and genomic organization of the histidine-rich protein of the malaria parasite Plasmodium lophurae. Nature. 1984 Dec 13;312(5995):616–620. doi: 10.1038/312616a0. [DOI] [PubMed] [Google Scholar]
  16. Rener J., Carter R., Rosenberg Y., Miller L. H. Anti-gamete monoclonal antibodies synergistically block transmission of malaria by preventing fertilization in the mosquito. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6797–6799. doi: 10.1073/pnas.77.11.6797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stahl H. D., Kemp D. J., Crewther P. E., Scanlon D. B., Woodrow G., Brown G. V., Bianco A. E., Anders R. F., Coppel R. L. Sequence of a cDNA encoding a small polymorphic histidine- and alanine-rich protein from Plasmodium falciparum. Nucleic Acids Res. 1985 Nov 11;13(21):7837–7846. doi: 10.1093/nar/13.21.7837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tokuyasu K. T. A technique for ultracryotomy of cell suspensions and tissues. J Cell Biol. 1973 May;57(2):551–565. doi: 10.1083/jcb.57.2.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Vernot-Hernandez J. P., Heidrich H. G. The relationship to knobs of the 92,000 D protein specific for knobby strains of Plasmodium falciparum. Z Parasitenkd. 1985;71(1):41–51. doi: 10.1007/BF00932917. [DOI] [PubMed] [Google Scholar]
  22. Vernot-Hernandez J. P., Heidrich H. G. Time-course of synthesis, transport and incorporation of a protein identified in purified membranes of host erythrocytes infected with a knob-forming strain of Plasmodium falciparum. Mol Biochem Parasitol. 1984 Jul;12(3):337–350. doi: 10.1016/0166-6851(84)90090-2. [DOI] [PubMed] [Google Scholar]

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