Intracellular signaling transduction in the differentiation of Trypanosoma cruzi: role of cAMP.

Abstract

We have studied the cell differentiation of Trypanosoma cruzi in an vitro system that allows the transformation of epimastigotes into metacyclic trypomastigotes. Intracellular cAMP levels of epimastigotes increased 3 fold prior to their differentiation into metacyclics where cAMP remained elevated 3.7 fold with respect to epimastigotes. We also observed a 3 fold increase in the specific activity of cAMP-binding of metacyclics crude homogenates. This activity resided in a cAMP-binding receptor protein (CARPT) which was different from the typical cAMP-binding subunits (RI and RII) of cAMP-dependent protein kinases, as shown by the use of polyclonal antibodies prepared against these two types of proteins. Anti-RI antibodies did not react with CARPT, and anti-RII antibodies gave a cross reaction with CARPT which was at least 1,000 fold less sensitive than the one shown by the homologous antigen. On Western blots CARPT displayed a major band with Mr = 87,000 instead of Mr = 56,000 for RII. These studies implicate that cAMP may act as a mediator of the cell differentiation of T. cruzi by a mechanism involving a novel type of cAMP-binding receptor.