Redox control in Neospora caninum: investigation of antioxidant system enzymes

• Grant number: 19/05758-5
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2019
• End date: October 31, 2020
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Ana Patricia Yatsuda Natsui
• Grantee: Jade Cabestre Venancio Brochi
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The apicomplexan protozoan Neospora caninum (Nc), the etiological agent of Neosporosis, is associated with neurological symptoms in dogs and abortion in cattle. This disease is worldwide spread and has no efficient treatment. The survival and replication of the parasite depends on an enzymatic antioxidant system defense for evading the oxidative stress inside the host cell. Among these enzymes, thioredoxin-dependent peroxide reductase (NcPrx) and glutathione reductase (NcGR) draw our attention due to a significant relative abundance in the proteome of Neospora caninum, allied to the absence of literature. Therefore, we will perform a characterization of the N. caninum antioxidant enzymes NcPrx and NcGR to elucidate their role in the successful parasite establishment and proliferation in the host cell. The NcGR and NcPrx recombinant proteins (rNcGR and rNcPrx) will be cloned and expressed followed by SDS-PAGE characterization and mass spectrometry identification. Through polyclonal antibodies production, NcGR and NcPrx will be detected in the parasite by immunofluorescence and western blot. The activity of the recombinant enzymes will be evaluated by specific enzymatic assays and their biochemical behavior against phenothiazine dyes and conoidin A will be tested. In addition, the action of NcPrx and NcGR in the response of N. caninum to oxidative stress will be demonstrated by the profile of antioxidant enzymes expressed after H2O2 stimulation and employing recombinant NcPrx and NcGR enzymes in tachyzoite proliferation assays in H2O2 presence. Ultimately, this project will reveal novel features of fundamental enzymes of the N. caninum redox system enabling investigations of drugs with unknown mechanisms of action and advances on the research of Neosporosis. (AU)

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)

VENANCIO-BROCHI, JADE CABESTRE; PEREIRA, LUIZ MIGUEL; BARONI, LUCIANA; ABREU-FILHO, PERICLES GAMA; YATSUDA, ANA PATRICIA. Characterization of the Neospora caninum peroxiredoxin: a novel peroxidase and antioxidant enzyme. Parasitology Research, v. 121, n. 6, p. 14-pg., 2022-04-01. (18/21020-3, 19/05758-5)

VENANCIO-BROCHI, JADE CABESTRE; PEREIRA, LUIZ MIGUEL; CALIL, FELIPE ANTUNES; TEIXEIRA, OLIVIA; BARONI, LUCIANA; ABREU-FILHO, PERICLES GAMA; BRAGA, GILBERTO UBIDA LEITE; NONATO, MARIA CRISTINA; YATSUDA, ANA PATRICIA. Glutathione reductase: A cytoplasmic antioxidant enzyme and a potential target for phenothiazinium dyes in Neospora caninum. International Journal of Biological Macromolecules, v. 187, p. 964-975, SEP 30 2021. (19/05758-5, 18/21020-3)

BROCHI, JADE CABESTRE VENANCIO; PEREIRA, LUIZ MIGUEL; YATSUDA, ANA PATRICIA. Extracellular H2O2, peroxiredoxin, and glutathione reductase alter Neospora caninum invasion and proliferation in Vero cells. Experimental Parasitology, v. 242, p. 6-pg., 2022-11-01. (19/05758-5)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

BROCHI, Jade Cabestre Venancio. Redox control in Neospora caninum: investigation of antioxidant system enzymes. 2022. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.