Proteins bearing cyclic nucleotide binding domains in Trypanosoma cruzi.

JOSE ESCALONA; MARTIN EDREIRA; GUILLERMO DI MARIO; GABRIEL FERRI

Mar Del Plata

Congreso; XXXI Reunion Anual Sociedad Argentina de Protozoologia; 2019

Cyclic AMP signaling have shown to be involved in Trypanosoma cruzi biology. However, little is now about the pathway in the parasite. Previous reports show that at the genomic level this parasite has several proteins that possess cyclic nucleotide binding domain (CNB), such as Protein kinase A. It has also been published that, unlike its mammalian counterpart, the trypanosomatid PKA does not bind cAMP and seems not to have the regulatory function of kinase activity. In order to increase the knowledge about the cAMP mediated pathway in T. cruzi, we have cloned and biochemically characterized cAMP binding proteins in T. cruzi. At the bioinformatic level, several proteins were found in the parasite?s genome. Six candidates, TcCLB.418221.20, TcCLB.504153.20, TcCLB.504449.30, TcCLB.508523.80, TcCLB.510691.30 and TcCLB.508273.30 were selected to cloned and fusion proteins to a his-tag and expressed in E. coli. Bacterially expressed proteins and a cAMP-agarose resin, were used in pull down and displacement experiments, in order to confirm cAMP binding. Interaction assays were then performed using trypomastigote lysates obtained from infected Vero cell cultures. The eluted proteins were analyzed by mass spectrometry obtaining potential partners candidates that will be futher studied.