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J. Biol. Chem., Vol. 276, Issue 50, 47078-47086, December 14, 2001

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Trypanosoma cruzi Prolyl Oligopeptidase Tc80 Is Involved in Nonphagocytic Mammalian Cell Invasion by Trypomastigotes^*

Philippe Grellier^§, Sandrine Vendeville^¶, Roger Joyeau, Izabela M. D. Bastos^**, Hervé Drobecq^¶, François Frappier, Antonio R. L. Teixeira^**, Joseph Schrével, Elisabeth Davioud-Charvet^¶, Christian Sergheraert^¶, and Jaime M. Santana^**

From the  Laboratoire de Biologie Parasitaire, Muséum National d'Histoire Naturelle, FR CNRS 63, 61 rue Buffon, 75231 Paris Cedex 05, France, the ^¶ Institut de Biologie de Lille-Institut Pasteur de Lille, UMR CNRS 8525, Faculté de Pharmacie, Université de Lille II, 1 rue Calmette, BP 447, 59024 Lille Cedex, France, the  Laboratoire de Chimie, Muséum National d'Histoire Naturelle, ESA CNRS 8041, 63 rue Buffon, 75231 Paris Cedex 05, France, and the ^** Laboratorio Multidisciplinar de Pesquisa em Doença de Chagas, Departamentos de Biologia Celular e de Patologia, Universidade de Brasilia, CP 04536, 70919-970, Brasilia, Distrito Federal, Brazil

Trypanosoma cruzi is an intracellular protozoan parasite able to invade a wide variety of mammalian cells. To have access to the target organs/cells, the parasite must cross the basal laminae and the extracellular matrix (ECM). We previously characterized an 80-kDa proteinase (Tc80) secreted by the infective trypomastigotes that hydrolyzes native collagens and might be involved in infection by degrading ECM components. Here, we present evidence indicating a role for Tc80 in the invasion of nonphagocytic cells. Tc80 was classified as a member of the prolyl oligopeptidase (POP) family of serine proteases and was also found to hydrolyze fibronectin. Selective inhibitors for POP Tc80 were synthesized that blocked parasite entry into cells. Blockage occurred when trypomastigotes were preincubated with irreversible inhibitors but not after host cell preincubation, and the blockage correlated with inhibition of POP Tc80 activity in treated parasites. These data and the enzyme location inside a vesicular compartment close to the flagellar pocket, a specialized domain in endocytosis/exocytosis, strongly suggest a role for POP Tc80 in the maturation of parasite protein(s) and/or, after secretion, in a local action on parasite or host cell/ECM components required for invasion.

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