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Characterization of enzymatic and lectin domains and the effects on innate immunity cell receptors

Grant number: 16/00629-4
Support Opportunities:Scholarships in Brazil - Post-Doctorate
Effective date (Start): July 01, 2016
Effective date (End): June 30, 2017
Field of knowledge:Biological Sciences - Immunology - Immunochemistry
Principal Investigator:Maria Cristina Roque Antunes Barreira
Grantee:Fabrício Freitas Fernandes
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:13/04088-0 - Lectin from pathogens, AP.TEM


Paracoccidioidomycosis (PCM), caused by the fungus thermally dimorphic P. brasiliensis, is a chronic systemic mycosis and granulomatous. The manifestations of the disease are associated with host and fungus factors, thus, the interaction between the fungal surface, and the molecules of the host immune system receptor lead to a kind of response can be beneficial or detrimental to the infected individual. For this reason, various studies are conducted to identify antigenic proteins of P. brasiliensis, which are important in host-pathogen interaction. The paracoccin is a candidate proteins under study. This protein is localized on the yeast surface, it was involved in fungal cell growth processes and virulence, and also has protective immune activity in experimental PCM. Given the importance of paracoccin activity in modulating an immune response and its role in fungal cell, it becomes mandatory to characterize the paracoccin domains and to determine the responsibility that each has on the activities of the entire molecule and identify its ligands. This project aims to clone, express and purify the truncated paracoccin in the lectin or enzymatic domains; identify the specificity of paracoccin recognition to different carbohydrates; identify the critical TLR4 N-glycosylation sites for recognition by paracoccin by means of point mutations in these N-glycosylation sites and expression in HEK cells; assess the participation of lectin and enzyme domains in modulating the immune response in vitro and in vivo; and verifying the paracoccin location in growth structures of the fungal cell, using the expression of proteins fused to GFP and DsRED through the ATMT technique. (AU)

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Scientific publications (4)
(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)
FERNANDES, FABRICIO F.; OLIVEIRA, ALINE F.; LANDGRAF, TAISE N.; CUNHA, CRISTINA; CARVALHO, AGOSTINHO; VENDRUSCOLO, PATRICIA E.; GONCALES, RELBER A.; ALMEIDA, FAUSTO; DA SILVA, THIAGO A.; RODRIGUES, FERNANDO; et al. Impact of Paracoccin Gene Silencing on Paracoccidioides brasiliensis Virulence. MBIO, v. 8, n. 4, . (14/05359-0, 14/22561-7, 16/04877-2, 13/04088-0, 12/09611-0, 16/00629-4)
GONCALES, RELBER AGUIAR; RICCI-AZEVEDO, RAFAEL; VIEIRA, VANESSA C. S.; FERNANDES, FABRICIO F.; THOMAZ, SANDRA M. DE O.; CARVALHO, AGOSTINHO; VENDRUSCOLO, PATRICIA E.; CUNHA, CRISTINA; ROQUE-BARREIRA, MARIA CRISTINA; RODRIGUES, FERNANDO. Paracoccin Overexpression in Paracoccidioides brasiliensis Enhances Fungal Virulence by Remodeling Chitin Properties of the Cell Wall. Journal of Infectious Diseases, v. 224, n. 1, p. 164-174, . (14/22561-7, 17/02998-0, 13/04088-0, 18/21708-5, 16/00629-4)
FABRÍCIO FREITAS FERNANDES; NAYLA S. PITANGUI; GABRIELA PERON; CLARISSA X. R. VALIM; VÍVIAN M. MAZUCATO; ALINE F. OLIVEIRA; THIAGO A. DA SILVA; MARIA-CRISTINA ROQUE-BARREIRA; ADEMILSON PANUNTO-CASTELO. Expression of Hsp60 and its cell location in Paracoccidioides brasiliensis. Revista do Instituto de Medicina Tropical de São Paulo, v. 62, . (16/00629-4, 17/01390-8)
LANDGRAF, TAISE N.; COSTA, MARCELO V.; OLIVEIRA, ALINE F.; RIBEIRO, WANDER C.; PANUNTO-CASTELO, ADEMILSON; FERNANDES, FABRICIO F.. Involvement of Dihydrolipoyl Dehydrogenase in the Phagocytosis and Killing of Paracoccidioides brasiliensis by Macrophages. FRONTIERS IN MICROBIOLOGY, v. 8, . (16/00629-4, 12/08552-0, 13/12278-3)

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