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. 1993 Jul;12(7):2881–2889. doi: 10.1002/j.1460-2075.1993.tb05950.x

Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum binds specifically to sulfated glycoconjugates and to HepG2 hepatoma cells suggesting a role for this molecule in sporozoite invasion of hepatocytes.

H M Müller 1, I Reckmann 1, M R Hollingdale 1, H Bujard 1, K J Robson 1, A Crisanti 1
PMCID: PMC413541  PMID: 8392935

Abstract

Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum contains an amino acid motif based around the sequence WSPCSVTCG which is also found in region II of the circumsporozoite (CS) proteins of different species of Plasmodium. This amino acid motif confers on the CS protein the ability to bind specifically to sulfated glycoconjugates and to hepatocytes. This suggests that the interaction of CS protein with sulfated glycoconjugates on the surface of the hepatocytes may represent the first molecular event of sporozoite invasion of liver cells. Experimental evidence indicates that TRAP is localized both on the micronemes and on the surface of P. falciparum sporozoites implying that TRAP with its putative sulfated glycoconjugate binding motif may also be involved in recognition and/or entry of hepatocytes by the sporozoite. We show here that different TRAP constructs expressed in Escherichia coli bind to sulfogalactosyl-cerebrosides (sulfatides) and to the surface of HepG2 cells. These interactions are dependent on the presence of the conserved amino acid motif WSPCSVTCG within the sequences of the constructs and are completely inhibited by several sulfated glycoconjugates as well as by suramin, a polysulfonated drug with anti-protozoan activity. Moreover, sporozoite invasion of HepG2 cells is inhibited by antisera raised against these different TRAP constructs and by the presence of low concentrations of suramin. We concluded that TRAP may be one of the parasite encoded molecules in the host-parasite interaction that results in sporozoite invasion of hepatocytes.

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Selected References

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

  1. Arnot D. E., Barnwell J. W., Tam J. P., Nussenzweig V., Nussenzweig R. S., Enea V. Circumsporozoite protein of Plasmodium vivax: gene cloning and characterization of the immunodominant epitope. Science. 1985 Nov 15;230(4727):815–818. doi: 10.1126/science.2414847. [DOI] [PubMed] [Google Scholar]
  2. Cerami C., Frevert U., Sinnis P., Takacs B., Clavijo P., Santos M. J., Nussenzweig V. The basolateral domain of the hepatocyte plasma membrane bears receptors for the circumsporozoite protein of Plasmodium falciparum sporozoites. Cell. 1992 Sep 18;70(6):1021–1033. doi: 10.1016/0092-8674(92)90251-7. [DOI] [PubMed] [Google Scholar]
  3. Cerami C., Kwakye-Berko F., Nussenzweig V. Binding of malarial circumsporozoite protein to sulfatides [Gal(3-SO4)beta 1-Cer] and cholesterol-3-sulfate and its dependence on disulfide bond formation between cysteines in region II. Mol Biochem Parasitol. 1992 Aug;54(1):1–12. doi: 10.1016/0166-6851(92)90089-3. [DOI] [PubMed] [Google Scholar]
  4. Clarke L. E., Tomley F. M., Wisher M. H., Foulds I. J., Boursnell M. E. Regions of an Eimeria tenella antigen contain sequences which are conserved in circumsporozoite proteins from Plasmodium spp. and which are related to the thrombospondin gene family. Mol Biochem Parasitol. 1990 Jun;41(2):269–279. doi: 10.1016/0166-6851(90)90190-w. [DOI] [PubMed] [Google Scholar]
  5. Cowan G., Krishna S., Crisanti A., Robson K. Expression of thrombospondin-related anonymous protein in Plasmodium falciparum sporozoites. Lancet. 1992 Jun 6;339(8806):1412–1413. doi: 10.1016/0140-6736(92)91229-2. [DOI] [PubMed] [Google Scholar]
  6. Dame J. B., Williams J. L., McCutchan T. F., Weber J. L., Wirtz R. A., Hockmeyer W. T., Maloy W. L., Haynes J. D., Schneider I., Roberts D. Structure of the gene encoding the immunodominant surface antigen on the sporozoite of the human malaria parasite Plasmodium falciparum. Science. 1984 Aug 10;225(4662):593–599. doi: 10.1126/science.6204383. [DOI] [PubMed] [Google Scholar]
  7. Dietrich C. P., Nader H. B., Straus A. H. Structural differences of heparan sulfates according to the tissue and species of origin. Biochem Biophys Res Commun. 1983 Mar 29;111(3):865–871. doi: 10.1016/0006-291x(83)91379-7. [DOI] [PubMed] [Google Scholar]
  8. Galinski M. R., Arnot D. E., Cochrane A. H., Barnwell J. W., Nussenzweig R. S., Enea V. The circumsporozoite gene of the Plasmodium cynomolgi complex. Cell. 1987 Jan 30;48(2):311–319. doi: 10.1016/0092-8674(87)90434-x. [DOI] [PubMed] [Google Scholar]
  9. Ginsburg V., Roberts D. D. Glycoconjugates and cell adhesion: the adhesive proteins laminin, thrombospondin and von Willebrand's factor bind specifically to sulfated glycolipids. Biochimie. 1988 Nov;70(11):1651–1659. doi: 10.1016/0300-9084(88)90300-8. [DOI] [PubMed] [Google Scholar]
  10. Goundis D., Reid K. B. Properdin, the terminal complement components, thrombospondin and the circumsporozoite protein of malaria parasites contain similar sequence motifs. Nature. 1988 Sep 1;335(6185):82–85. doi: 10.1038/335082a0. [DOI] [PubMed] [Google Scholar]
  11. Hedstrom R. C., Campbell J. R., Leef M. L., Charoenvit Y., Carter M., Sedegah M., Beaudoin R. L., Hoffman S. L. A malaria sporozoite surface antigen distinct from the circumsporozoite protein. Bull World Health Organ. 1990;68 (Suppl):152–157. [PMC free article] [PubMed] [Google Scholar]
  12. Hollingdale M. R., Nardin E. H., Tharavanij S., Schwartz A. L., Nussenzweig R. S. Inhibition of entry of Plasmodium falciparum and P. vivax sporozoites into cultured cells; an in vitro assay of protective antibodies. J Immunol. 1984 Feb;132(2):909–913. [PubMed] [Google Scholar]
  13. Holt G. D., Krivan H. C., Gasic G. J., Ginsburg V. Antistasin, an inhibitor of coagulation and metastasis, binds to sulfatide (Gal(3-SO4) beta 1-1Cer) and has a sequence homology with other proteins that bind sulfated glycoconjugates. J Biol Chem. 1989 Jul 25;264(21):12138–12140. [PubMed] [Google Scholar]
  14. Holt G. D., Pangburn M. K., Ginsburg V. Properdin binds to sulfatide [Gal(3-SO4)beta 1-1 Cer] and has a sequence homology with other proteins that bind sulfated glycoconjugates. J Biol Chem. 1990 Feb 15;265(5):2852–2855. [PubMed] [Google Scholar]
  15. Klar A., Baldassare M., Jessell T. M. F-spondin: a gene expressed at high levels in the floor plate encodes a secreted protein that promotes neural cell adhesion and neurite extension. Cell. 1992 Apr 3;69(1):95–110. doi: 10.1016/0092-8674(92)90121-r. [DOI] [PubMed] [Google Scholar]
  16. Krivan H. C., Olson L. D., Barile M. F., Ginsburg V., Roberts D. D. Adhesion of Mycoplasma pneumoniae to sulfated glycolipids and inhibition by dextran sulfate. J Biol Chem. 1989 Jun 5;264(16):9283–9288. [PubMed] [Google Scholar]
  17. Lawler J., Hynes R. O. The structure of human thrombospondin, an adhesive glycoprotein with multiple calcium-binding sites and homologies with several different proteins. J Cell Biol. 1986 Nov;103(5):1635–1648. doi: 10.1083/jcb.103.5.1635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leung-Hagesteijn C., Spence A. M., Stern B. D., Zhou Y., Su M. W., Hedgecock E. M., Culotti J. G. UNC-5, a transmembrane protein with immunoglobulin and thrombospondin type 1 domains, guides cell and pioneer axon migrations in C. elegans. Cell. 1992 Oct 16;71(2):289–299. doi: 10.1016/0092-8674(92)90357-i. [DOI] [PubMed] [Google Scholar]
  19. Ortega-Barria E., Pereira M. E. A novel T. cruzi heparin-binding protein promotes fibroblast adhesion and penetration of engineered bacteria and trypanosomes into mammalian cells. Cell. 1991 Oct 18;67(2):411–421. doi: 10.1016/0092-8674(91)90192-2. [DOI] [PubMed] [Google Scholar]
  20. Pancake S. J., Holt G. D., Mellouk S., Hoffman S. L. Malaria sporozoites and circumsporozoite proteins bind specifically to sulfated glycoconjugates. J Cell Biol. 1992 Jun;117(6):1351–1357. doi: 10.1083/jcb.117.6.1351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Petitou M., Lormeau J. C., Choay J. Interaction of heparin and antithrombin III. The role of O-sulfate groups. Eur J Biochem. 1988 Oct 1;176(3):637–640. doi: 10.1111/j.1432-1033.1988.tb14324.x. [DOI] [PubMed] [Google Scholar]
  22. Pierschbacher M. D., Ruoslahti E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984 May 3;309(5963):30–33. doi: 10.1038/309030a0. [DOI] [PubMed] [Google Scholar]
  23. Prater C. A., Plotkin J., Jaye D., Frazier W. A. The properdin-like type I repeats of human thrombospondin contain a cell attachment site. J Cell Biol. 1991 Mar;112(5):1031–1040. doi: 10.1083/jcb.112.5.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roberts D. D., Haverstick D. M., Dixit V. M., Frazier W. A., Santoro S. A., Ginsburg V. The platelet glycoprotein thrombospondin binds specifically to sulfated glycolipids. J Biol Chem. 1985 Aug 5;260(16):9405–9411. [PubMed] [Google Scholar]
  25. Roberts D. D. Interactions of thrombospondin with sulfated glycolipids and proteoglycans of human melanoma cells. Cancer Res. 1988 Dec 1;48(23):6785–6793. [PubMed] [Google Scholar]
  26. Robson K. J., Hall J. R., Davies L. C., Crisanti A., Hill A. V., Wellems T. E. Polymorphism of the TRAP gene of Plasmodium falciparum. Proc Biol Sci. 1990 Dec 22;242(1305):205–216. doi: 10.1098/rspb.1990.0126. [DOI] [PubMed] [Google Scholar]
  27. Robson K. J., Hall J. R., Jennings M. W., Harris T. J., Marsh K., Newbold C. I., Tate V. E., Weatherall D. J. A highly conserved amino-acid sequence in thrombospondin, properdin and in proteins from sporozoites and blood stages of a human malaria parasite. Nature. 1988 Sep 1;335(6185):79–82. doi: 10.1038/335079a0. [DOI] [PubMed] [Google Scholar]
  28. Rogers W. O., Malik A., Mellouk S., Nakamura K., Rogers M. D., Szarfman A., Gordon D. M., Nussler A. K., Aikawa M., Hoffman S. L. Characterization of Plasmodium falciparum sporozoite surface protein 2. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9176–9180. doi: 10.1073/pnas.89.19.9176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  30. Sun X., Mosher D. F., Rapraeger A. Heparan sulfate-mediated binding of epithelial cell surface proteoglycan to thrombospondin. J Biol Chem. 1989 Feb 15;264(5):2885–2889. [PubMed] [Google Scholar]
  31. 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]
  32. Weber J. L., Egan J. E., Lyon J. A., Wirtz R. A., Charoenvit Y., Maloy W. L., Hockmeyer W. T. Plasmodium berghei: cloning of the circumsporozoite protein gene. Exp Parasitol. 1987 Jun;63(3):295–300. doi: 10.1016/0014-4894(87)90176-7. [DOI] [PubMed] [Google Scholar]
  33. Zabrenetzky V. S., Kohn E. C., Roberts D. D. Suramin inhibits laminin- and thrombospondin-mediated melanoma cell adhesion and migration and binding of these adhesive proteins to sulfatide. Cancer Res. 1990 Sep 15;50(18):5937–5942. [PubMed] [Google Scholar]
  34. Zhang J. P., Stephens R. S. Mechanism of C. trachomatis attachment to eukaryotic host cells. Cell. 1992 May 29;69(5):861–869. doi: 10.1016/0092-8674(92)90296-o. [DOI] [PubMed] [Google Scholar]
  35. de la Cruz V. F., Lal A. A., McCutchan T. F. Sequence variation in putative functional domains of the circumsporozoite protein of Plasmodium falciparum. Implications for vaccine development. J Biol Chem. 1987 Sep 5;262(25):11935–11939. [PubMed] [Google Scholar]

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