This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Manque, P. M.
Right arrow Articles by Yoshida, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Manque, P. M.
Right arrow Articles by Yoshida, N.

 Previous Article  |  Next Article 

Infection and Immunity, February 2000, p. 478-484, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of the Cell Adhesion Site of Trypanosoma cruzi Metacyclic Stage Surface Glycoprotein gp82

Patricio M. Manque,1 Daniel Eichinger,2 Maria A. Juliano,3 Luiz Juliano,3 Jorge E. Araya,1,dagger and Nobuko Yoshida1,*

Departamento de Microbiologia, Imunologia e Parasitologia,1 and Departamento de Biofisica,3 Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil, and Department of Medical and Molecular Parasitology, New York University School of Medicine, New York, New York2

Received 12 August 1999/Returned for modification 7 October 1999/Accepted 9 November 1999

The surface glycoprotein gp82, expressed in the insect-stage metacyclic trypomastigotes of Trypanosoma cruzi, has been implicated in mammalian cell invasion. Here we have characterized the cell adhesion site of gp82 by using recombinant proteins and synthetic peptides based on gp82. The recombinant protein Del-4/8, lacking 65 amino acids of gp82 central domain (at positions 257 to 321), was virtually devoid of cell-binding activity and lacked the ability to inhibit parasite invasion, in contrast to J18, the construct containing the full-length gp82 sequence (amino acids 1 to 516). Constructs with shorter deletions, i.e., Del-4 (deleted from 257 to 271) and Del-8 (deleted from 293 to 321), bound to target cells to a significantly lesser degree than did J18. The sites deleted in recombinant proteins Del-4 and Del-8 contained acidic amino acids critical for cell adhesion. Thus, the cell-binding capacity of protein Del-E/D, lacking the glutamic acid (259/260) and aspartic acid (303/304) pairs, was negligible, as was its capacity to inhibit parasite internalization. Of a set of synthetic peptides spanning the gp82 central domain, a 22-mer hybrid peptide, p4/8, formed by two noncontiguous sequences (at positions 257 to 273 and 302 to 306) and containing the four acidic residues, competed with the binding of J18 protein to target cells and significantly inhibited (~60%) the penetration of parasites. This peptide, generated by the juxtaposition of sequences that are separated by a hydrophobic stretch in the linear molecule, appears to be mimicking a conformation-dependent cell-binding site of gp82. Experiments of antibody competition with a set of 20-mer overlapping peptides mapped the epitope for 3F6, a monoclonal antibody directed to gp82 that inhibits parasite invasion, to the sequence represented by peptide p3 (244 to 263), which has a partial overlap with the cell adhesion site.

* Corresponding author. Mailing address: Escola Paulista de Medicina, Universidade Federal de São Paulo, R. Botucatu, 862- 6° andar, 04023-062, São Paulo, S.P., Brazil. Phone: 55-11-576-4532. Fax: 55-11-571-1095. E-mail: nyoshida.dmip{at}epm.br.

dagger Present address: Universidad de Antofagasta, Antofagasta, Chile.

Infection and Immunity, February 2000, p. 478-484, Vol. 68, No. 2
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Atayde, V. D., Cortez, M., Souza, R., da Silveira, J. F., Yoshida, N. (2007). Expression and Cellular Localization of Molecules of the gp82 Family in Trypanosoma cruzi Metacyclic Trypomastigotes. Infect. Immun. 75: 3264-3270 [Abstract] [Full Text]  
  • Ferreira, D., Cortez, M., Atayde, V. D., Yoshida, N. (2006). Actin Cytoskeleton-Dependent and -Independent Host Cell Invasion by Trypanosoma cruzi Is Mediated by Distinct Parasite Surface Molecules.. Infect. Immun. 74: 5522-5528 [Abstract] [Full Text]  
  • Cortez, M., Neira, I., Ferreira, D., Luquetti, A. O., Rassi, A., Atayde, V. D., Yoshida, N. (2003). Infection by Trypanosoma cruzi Metacyclic Forms Deficient in gp82 but Expressing a Related Surface Molecule, gp30. Infect. Immun. 71: 6184-6191 [Abstract] [Full Text]  
  • Manque, P. M., Neira, I., Atayde, V. D., Cordero, E., Ferreira, A. T., da Silveira, J. F., Ramirez, M., Yoshida, N. (2003). Cell Adhesion and Ca2+ Signaling Activity in Stably Transfected Trypanosoma cruzi Epimastigotes Expressing the Metacyclic Stage-Specific Surface Molecule gp82. Infect. Immun. 71: 1561-1565 [Abstract] [Full Text]  
  • Neira, I., Silva, F. A., Cortez, M., Yoshida, N. (2003). Involvement of Trypanosoma cruzi Metacyclic Trypomastigote Surface Molecule gp82 in Adhesion to Gastric Mucin and Invasion of Epithelial Cells. Infect. Immun. 71: 557-561 [Abstract] [Full Text]  
  • Malaga, S., Yoshida, N. (2001). Targeted Reduction in Expression of Trypanosoma cruzi Surface Glycoprotein gp90 Increases Parasite Infectivity. Infect. Immun. 69: 353-359 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»