Methods to Investigate Signal Transduction Pathways in Trypanosoma cruzi: Cyclic Nucleotide Phosphodiesterases Assay Protocols

SCHOIJET, ALEJANDRA C.; STERNLIEB, TAMARA; ALONSO, GUILLERMO D.

METHODS IN MOLECULAR BIOLOGY (CLIFTON, N.J.)

Springer Nature

Lugar: Picassoplatz; Año: 2020 vol. 2116 p. 523 - 534

1064-3745

Intracellular levels of cyclic nucleotide second messengers are regulated predominantly by a large superfamily of phosphodiesterases (PDEs). Most of the different PDE variants play specific physiological functions; in fact, PDEs can associate with other proteins allowing them to be strategically anchored throughout the cell. In this regard, precise cellular expression and compartmentalization of these enzymes produce the specific control of cAMP and cGMP gradients in cells and enable their integration with other signaling pathways.In trypanosomatids, some PDEs are essential for their survival and play fundamental roles in the adaptation of these parasites to different environmental stresses, as well as in the differentiation between their different life cycle forms. Given that these enzymes are similar to human PDEs, but also have differential biochemical properties, and due to the great knowledge of drugs that target human PDEs, trypanosomatid PDEs could be postulated as important therapeutic targets through the repositioning of drugs.In this chapter we describe a simple and sensitive radioisotope-based method to measure cyclic 3´,5´-nucleotide phosphodiesterase using [3H]cAMP.