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Infection and Immunity, October 2003, p. 5739-5749, Vol. 71, No. 10
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.10.5739-5749.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

gp63 Homologues in Trypanosoma cruzi: Surface Antigens with Metalloprotease Activity and a Possible Role in Host Cell Infection

Ileana C. Cuevas, Juan J. Cazzulo, and Daniel O. Sánchez^*

Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, Universidad Nacional de General San Martín, Consejo Nacional de Investigaciones Científicas y Técnicas, San Martín, Provincia de Buenos Aires, Argentina

Received 14 April 2003/ Returned for modification 29 May 2003/ Accepted 26 July 2003

gp63 is a highly abundant glycosylphosphatidylinositol (GPI)-anchored membrane protein expressed predominantly in the promastigote but also in the amastigote stage of Leishmania species. In Leishmania spp., gp63 has been implicated in a number of steps in establishment of infection. Here we demonstrate that Trypanosoma cruzi, the etiological agent of Chagas' disease, has a family of gp63 genes composed of multiple groups. Two of these groups, Tcgp63-I and -II, are present as high-copy-number genes. The genomic organization and mRNA expression pattern were specific for each group. Tcgp63-I was widely expressed, while the Tcgp63-II group was scarcely detected in Northern blots, even though it is well represented in the T. cruzi genome. Western blots using sera directed against a synthetic peptide indicated that the Tcgp63-I group produced proteins of 78 kDa, differentially expressed during the life cycle. Immunofluorescence staining and phosphatidylinositol-specific phospholipase C digestion confirmed that Tcgp63-I group members are surface proteins bound to the membrane by a GPI anchor. We also demonstrate the presence of metalloprotease activity which is attributable, at least in part, to Tcgp63-I group. Since antibodies against Tcgp63-I partially blocked infection of Vero cells by trypomastigotes, a possible role for this group in infection is suggested.

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* Corresponding author. Mailing address: Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín, INTI, Edificio 24, 1650, San Martin, Buenos Aires, Argentina. Phone: 54-11-4580-7255. Fax: 54-11-4752-9639. E-mail: dsanchez{at}iib.unsam.edu.ar.

Editor: W. A. Petri, Jr.

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Infection and Immunity, October 2003, p. 5739-5749, Vol. 71, No. 10
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.10.5739-5749.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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