Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1993 Oct;61(10):4196–4201. doi: 10.1128/iai.61.10.4196-4201.1993

Characterization of a cDNA clone encoding the carboxy-terminal domain of a 90-kilodalton surface antigen of Trypanosoma cruzi metacyclic trypomastigotes.

F R Franco 1, G S Paranhos-Bacallà 1, L M Yamauchi 1, N Yoshida 1, J F da Silveira 1
PMCID: PMC281144  PMID: 8406808

Abstract

We have cloned and sequenced a cDNA for a metacyclic trypomastigote-specific glycoprotein with a molecular mass of 90 kDa, termed MTS-gp90. By immunoblotting, antibodies to the MTS-gp90 recombinant protein reacted exclusively with a 90-kDa antigen of metacyclic trypomastigotes. The insert of the MTS-gp90 cDNA clone strongly hybridized with a single 3.0-kb mRNA of metacyclic forms, whereas the hybridization signal with epimastigote mRNA was weak and those with RNAs from other developmental stages were negative, indicating that transcription of the MTS-gp90 gene is developmentally regulated. A series of experiments showed that the MTS-gp90 gene is present in multiple copies in the Trypanosoma cruzi genome, arranged in a nontandem manner, and that there are at least 40 copies of the gene per haploid genome. Sequence analysis of recombinant MTS-gp90 revealed 40 to 60% identity at the amino acid level with members of a family of mammalian stage-specific, 85-kDa surface antigens of T. cruzi. However, there are considerable differences in the amino acid compositions outside the homology region.

Full text

PDF
4196

Images in this article

4197Image
on p.4197
4197Image
on p.4197
4197Image
on p.4197
4199Image
on p.4199
4200Image
on p.4200

Selected References

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

  1. Affranchino J. L., Ibañez C. F., Luquetti A. O., Rassi A., Reyes M. B., Macina R. A., Aslund L., Pettersson U., Frasch A. C. Identification of a Trypanosoma cruzi antigen that is shed during the acute phase of Chagas' disease. Mol Biochem Parasitol. 1989 May 15;34(3):221–228. doi: 10.1016/0166-6851(89)90050-9. [DOI] [PubMed] [Google Scholar]
  2. Alves M. J., Abuin G., Kuwajima V. Y., Colli W. Partial inhibition of trypomastigote entry into cultured mammalian cells by monoclonal antibodies against a surface glycoprotein of Trypanosoma cruzi. Mol Biochem Parasitol. 1986 Oct;21(1):75–82. doi: 10.1016/0166-6851(86)90081-2. [DOI] [PubMed] [Google Scholar]
  3. Araguth M. F., Rodrigues M. M., Yoshida N. Trypanosoma cruzi metacyclic trypomastigotes: neutralization by the stage-specific monoclonal antibody 1G7 and immunogenicity of 90 kD surface antigen. Parasite Immunol. 1988 Nov;10(6):707–712. doi: 10.1111/j.1365-3024.1988.tb00256.x. [DOI] [PubMed] [Google Scholar]
  4. Boschetti M. A., Piras M. M., Henríquez D., Piras R. The interaction of a Trypanosoma cruzi surface protein with Vero cells and its relationship with parasite adhesion. Mol Biochem Parasitol. 1987 Jun;24(2):175–184. doi: 10.1016/0166-6851(87)90104-6. [DOI] [PubMed] [Google Scholar]
  5. Campetella O., Sánchez D., Cazzulo J. J., Frasch A. C. A superfamily of Trypanosoma cruzi surface antigens. Parasitol Today. 1992 Nov;8(11):378–381. doi: 10.1016/0169-4758(92)90175-2. [DOI] [PubMed] [Google Scholar]
  6. Cotrim P. C., Paranhos G. S., Mortara R. A., Wanderley J., Rassi A., Camargo M. E., da Silveira J. F. Expression in Escherichia coli of a dominant immunogen of Trypanosoma cruzi recognized by human chagasic sera. J Clin Microbiol. 1990 Mar;28(3):519–524. doi: 10.1128/jcm.28.3.519-524.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cross G. A. Glycolipid anchoring of plasma membrane proteins. Annu Rev Cell Biol. 1990;6:1–39. doi: 10.1146/annurev.cb.06.110190.000245. [DOI] [PubMed] [Google Scholar]
  8. Davis C. D., Kuhn R. E. Selective binding of Trypanosoma cruzi to host cell membrane polypeptides. Infect Immun. 1990 Jan;58(1):1–6. doi: 10.1128/iai.58.1.1-6.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fouts D. L., Ruef B. J., Ridley P. T., Wrightsman R. A., Peterson D. S., Manning J. E. Nucleotide sequence and transcription of a trypomastigote surface antigen gene of Trypanosoma cruzi. Mol Biochem Parasitol. 1991 Jun;46(2):189–200. doi: 10.1016/0166-6851(91)90043-6. [DOI] [PubMed] [Google Scholar]
  11. González J., Araguth M. F., Yoshida N. Resistance to acute Trypanosoma cruzi infection resulting from immunization of mice with a 90-kilodalton antigen from metacyclic trypomastigotes. Infect Immun. 1991 Mar;59(3):863–867. doi: 10.1128/iai.59.3.863-867.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Güther M. L., de Almeida M. L., Yoshida N., Ferguson M. A. Structural studies on the glycosylphosphatidylinositol membrane anchor of Trypanosoma cruzi 1G7-antigen. The structure of the glycan core. J Biol Chem. 1992 Apr 5;267(10):6820–6828. [PubMed] [Google Scholar]
  13. Kahn S., Van Voorhis W. C., Eisen H. The major 85-kD surface antigen of the mammalian form of Trypanosoma cruzi is encoded by a large heterogeneous family of simultaneously expressed genes. J Exp Med. 1990 Aug 1;172(2):589–597. doi: 10.1084/jem.172.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Martins M. S., Hudson L., Krettli A. U., Cançado J. R., Brener Z. Human and mouse sera recognize the same polypeptide associated with immunological resistance to Trypanosoma cruzi infection. Clin Exp Immunol. 1985 Aug;61(2):343–350. [PMC free article] [PubMed] [Google Scholar]
  16. Mortara R. A., Araguth M. F., Yoshida N. Reactivity of stage-specific monoclonal antibody 1G7 with metacyclic trypomastigotes of Trypanosoma cruzi strains: lytic property and 90,000 mol. wt surface antigen polymorphism. Parasite Immunol. 1988 Jul;10(4):369–378. doi: 10.1111/j.1365-3024.1988.tb00227.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Ozaki L. S., Mattei D., Jendoubi M., Druihle P., Blisnick T., Guillotte M., Puijalon O., Da Silva L. P. Plaque antibody selection: rapid immunological analysis of a large number of recombinant phage clones positive to sera raised against Plasmodium falciparum antigens. J Immunol Methods. 1986 May 22;89(2):213–219. doi: 10.1016/0022-1759(86)90360-1. [DOI] [PubMed] [Google Scholar]
  19. Pereira M. E., Mejia J. S., Ortega-Barria E., Matzilevich D., Prioli R. P. The Trypanosoma cruzi neuraminidase contains sequences similar to bacterial neuraminidases, YWTD repeats of the low density lipoprotein receptor, and type III modules of fibronectin. J Exp Med. 1991 Jul 1;174(1):179–191. doi: 10.1084/jem.174.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Peterson D. S., Fouts D. L., Manning J. E. The 85-kd surface antigen gene of Trypanosoma cruzi is telomeric and a member of a multigene family. EMBO J. 1989 Dec 1;8(12):3911–3916. doi: 10.1002/j.1460-2075.1989.tb08571.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Peterson D. S., Wrightsman R. A., Manning J. E. Cloning of a major surface-antigen gene of Trypanosoma cruzi and identification of a nonapeptide repeat. Nature. 1986 Aug 7;322(6079):566–568. doi: 10.1038/322566a0. [DOI] [PubMed] [Google Scholar]
  22. Ruiz R. de C., Rigoni V. L., Gonzalez J., Yoshida N. The 35/50 kDa surface antigen of Trypanosoma cruzi metacyclic trypomastigotes, an adhesion molecule involved in host cell invasion. Parasite Immunol. 1993 Feb;15(2):121–125. doi: 10.1111/j.1365-3024.1993.tb00591.x. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schenkman S., Yoshida N., Cardoso de Almeida M. L. Glycophosphatidylinositol-anchored proteins in metacyclic trypomastigotes of Trypanosoma cruzi. Mol Biochem Parasitol. 1988 Jun;29(2-3):141–151. doi: 10.1016/0166-6851(88)90069-2. [DOI] [PubMed] [Google Scholar]
  25. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  26. Takle G. B., Cross G. A. An 85-kilodalton surface antigen gene family of Trypanosoma cruzi encodes polypeptides homologous to bacterial neuraminidases. Mol Biochem Parasitol. 1991 Oct;48(2):185–198. doi: 10.1016/0166-6851(91)90114-l. [DOI] [PubMed] [Google Scholar]
  27. Takle G. B., Young A., Snary D., Hudson L., Nicholls S. C. Cloning and expression of a trypomastigote-specific 85-kilodalton surface antigen gene from Trypanosoma cruzi. Mol Biochem Parasitol. 1989 Nov;37(1):57–64. doi: 10.1016/0166-6851(89)90102-3. [DOI] [PubMed] [Google Scholar]
  28. Teixeira M. M., Yoshida N. Stage-specific surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi identified by monoclonal antibodies. Mol Biochem Parasitol. 1986 Mar;18(3):271–282. doi: 10.1016/0166-6851(86)90085-x. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Yoshida N., Mortara R. A., Araguth M. F., Gonzalez J. C., Russo M. Metacyclic neutralizing effect of monoclonal antibody 10D8 directed to the 35- and 50-kilodalton surface glycoconjugates of Trypanosoma cruzi. Infect Immun. 1989 Jun;57(6):1663–1667. doi: 10.1128/iai.57.6.1663-1667.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yoshida N. Surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi. Infect Immun. 1983 May;40(2):836–839. doi: 10.1128/iai.40.2.836-839.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES