Scholarship 18/03727-2 - Trypanosoma cruzi, Transdução de sinais - BV FAPESP
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The role of second messengers in the signaling pathways of the Trypanosoma Cruzi interaction with the extracellular matrix

Grant number: 18/03727-2
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Start date until: June 01, 2018
End date until: May 31, 2021
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Principal Investigator:Maria Julia Manso Alves
Grantee:Nubia Carolina Manchola Varón
Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Post-translational modifications by phosphorylation and s-nitrosylation were detected by proteomic analysis in several key enzymes of cell signaling processes in the parasite when it comes into contact with ECM. Some of these proteins are involved in cAMP, cGMP and calcium-mediated signaling processes, such as cAMP phosphodiesterase (PDE) and calmodulin. This project aims to validate the data obtained by proteomic analysis and to study whether there is a relationship between the presence or absence of calcium and the appearance of some of these modifications on contact with ECM. Changes in the calcium turnover profile will be monitored using Fluo-4 fluorophore in the presence of ECM and BAPTA/AM (a calcium chelator). Signaling triggered will be accompanied by quantification of cAMP and cGMP produced under the different conditions. We will make comparisons between changes in the profiles of phosphorylation and S-nitrosylation modifications between parasites incubated with ECM and BAPTA/AM, verifying whether these modifications are present or absent in proteins of interest in some related signaling pathways such as PKA and MAPK, which can be verified with antibodies specific for phosphorylation, or immuno-precipitated and analyzed for S-nitrosylation and phosphorylation. The enzymatic activities of nitrate synthase an enzyme involved in the production of nitric oxide in the parasite and PDE, a regulatory enzyme of the intracellular levels of cAMP, will be evaluated from total extracts of parasites previously incubated with ECM. The aim of this project is to increase knowledge under the signaling processes that occur between trypomastigote forms and ECM, a key interaction for the internalization of T. cruzi in cells. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MATTOS, ELICIANE C.; CANUTO, GISELE; MANCHOLA, NUBIA C.; MAGALHAES, RUBENS D. M.; CROZIER, THOMAS W. M.; LAMONT, DOUGLAS J.; TAVARES, MARINA F. M.; COLLI, WALTER; FERGUSON, MICHAEL A. J.; ALVES, MARIA JULIA M.. Reprogramming of Trypanosoma cruzi metabolism triggered by parasite interaction with the host cell extracellular matrix. PLoS Neglected Tropical Diseases, v. 13, n. 2, . (16/14506-1, 14/25494-9, 18/03727-2, 13/16002-2)
MULE, S. N.; MANCHOLA, N. C.; DE OLIVEIRA, G. S.; PEREIRA, M.; MAGALHAES, R. D. M.; TEIXEIRA, A. A.; COLLI, W.; ALVES, M. J. M.; PALMISANO, G.. Proteome-wide modulation of S-nitrosylation in Trypanosoma cruzi trypomastigotes upon interaction with the host extracellular matrix. JOURNAL OF PROTEOMICS, v. 231, . (14/25494-9, 14/10046-0, 18/03727-2, 17/04032-5, 18/18257-1, 18/13283-4, 18/15549-1, 12/03887-3)

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