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Originally published In Press as doi:10.1074/jbc.M000253200 on June 7, 2000

J. Biol. Chem., Vol. 275, Issue 36, 27671-27680, September 8, 2000
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Trypanosoma cruzi Surface Mucins with Exposed Variant Epitopes*

Guido D. PollevickDagger §, Javier M. Di NoiaDagger §||, Maria L. Salto**Dagger Dagger , Carlos Lima**§§, M. Susana Leguizamón¶¶, Rosa M. de Lederkremer**, and Alberto C. C. FraschDagger ||||

From the Dagger  Instituto de Investigaciones Biotecnológicas, Instituto Tecnológico de Chascomús (CONICET), Universidad Nacional de General San Martín, Av. Gral. Paz s/n, INTI, Edificio 24, 1650, San Martín, Pcia. de Buenos Aires, ** CIHIDECAR (CONICET) Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, and the ¶¶ Departamento de Microbiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

The protozoan parasite Trypanosoma cruzi, the agent of Chagas disease, has a large number of mucin molecules on its surface, whose expression is regulated during the life cycle. These mucins are the main acceptors of sialic acid, a monosaccharide that is required by the parasite to infect and survive in the mammalian host. A large mucin-like gene family named TcMUC containing about 500 members has been identified previously in T. cruzi. TcMUC can be divided into two subfamilies according to the presence or absence of tandem repeats in the central region of the genes. In this work, T. cruzi parasites were transfected with one tagged member of each subfamily. Only the product from the gene with repeats was highly O-glycosylated in vivo. The O-linked oligosaccharides consisted mainly of beta -D-Galp(1right-arrow4)GlcNAc and beta -D-Galp(1right-arrow4)[beta -D-Galp(1right-arrow6)]-D-GlcNAc. The same glycosyl moieties were found in endogenous mucins. The mature product was anchored by glycosylphosphatidylinositol to the plasma membrane and exposed to the medium. Sera from infected mice recognized the recombinant product of one repeats-containing gene thus showing that they are expressed during the infection. TcMUC genes encode a hypervariable region at the N terminus. We now show that the hypervariable region is indeed present in the exposed mature N termini of the mucins because sera from infected hosts recognized peptides having sequences from this region. The results are discussed in comparison with the mucins from the insect stages of the parasite (Di Noia, J. M., D'Orso, I., Sánchez, D. O., and Frasch, A. C. C. (2000) J. Biol. Chem. 275, 10218-10227) which do not have variable regions.

* This work was supported in part by grants from the Agencia Nacional de Promoción Científica y Tecnológica, Argentina, the Swedish Agency for Research Cooperation with Developing Countries (SAREC-SIDA), the International Atomic Energy Agency, the Consejo Nacional de Investigaciones Científicas y Técnicas, and the University of Buenos Aires.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ The first two authors contributed equally to this work.

CONICET researchers.

|| CONICET fellow.

Dagger Dagger University of Buenos Aires fellow.

§§ CONICET principal professional.

|||| Supported in part by an International Research Scholar grant from the Howard Hughes Medical Institute. To whom correspondence should be addressed. Tel.: 54-11-4580-7255; Fax: 54-11-4752-9639; E-mail: cfrasch@iib.unsam.edu.ar.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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