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Inhibitor and proteases of ectoparasites: relationship of structure-function and identification of the role of these molecules in the interaction of diseases vector e their etiological agents

Grant number: 12/03657-8
Support type:Research Projects - Thematic Grants
Duration: October 01, 2012 - September 30, 2018
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Aparecida Sadae Tanaka
Grantee:Aparecida Sadae Tanaka
Home Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Assoc. researchers:Andréa Cristina Fogaça ; Anita Mitico Tanaka-Azevedo ; Francisco Jose Alves Lemos ; Itabajara da Silva Vaz Junior ; Marcos Horacio Pereira ; Margareth de Lara Capurro-Guimarães ; Mariana da Silva Araujo ; Ricardo Nascimento Araujo ; Sergio Schenkman ; Sirlei Daffre
Associated grant(s):17/50409-3 - Searching for proteases and their inhibitors in ticks to be used in vaccine development, AP.R SPRINT
Associated scholarship(s):16/09874-1 - Functional studies of protease inhibitors using tick and mosquito cell cultures, BP.PD
16/03736-6 - Cloning, expression and purification of a chimeric protein based on Rhipicephalus microplus proteins, BP.IC
15/09268-1 - Characterization of two proteases from Babesia bovis parasite: Importance in the host infections., BP.DR
+ associated scholarships 14/22464-1 - Cloning, expression and purification of an aspartyl protease (Rm27441) presents in the gut of Rhipicephalus microplus tick, BP.IC
12/20597-9 - Interaction of proteases from Babesia bovis, the etiological agent of cattle babebiosis, with protease inhibitors from vector tick Rhiphicephallus (Boophilus) microplus, and its potential for vaccination., BP.PD
13/01175-9 - Functional study of protease BmSEI present in the gut of the tick Rhipicephalus microplus and their participation in infection with Anaplasma marginale., BP.DR
12/15191-3 - Functional characterization of serine protease inhibitors present in mosquito Aedes aegypti infected and not infected with dengue virus, BP.PD - associated scholarships

Abstract

The knowledge of the diversity of molecules produced by disease vectors in response to their etiologic agents is an important strategy to identify important targets in these vectors physiology, which could be used in rational design of drugs and pesticides. In attempt to contribute in this research field, this project has as general objective understand the role of protease inhibitors, proteolytic enzymes and hypothetical proteins identified in previous projects (using the following models of study: Triatoma infestans, one of the main vectors of Chagas Disease in Latin America, Aedes aegypti mosquito, vector of Dengue and the tick species Rhipicephalus (Boophilus) microplus, ectoparasite of cattle) in the interaction of these vectors with its etiological agents (Trypanosoma cruzi, dengue virus and Anaplasma marginale). The experience of the group in the research related to structure and function of protease inhibitors and anticoagulants (of the three vectors used in this study) will facilitate the implementation of the proposed project. To ensure the success of this project, the experiments of infection of the vector with their etiological agents will be carried out in collaboration with researchers who already have proven experience in the models of infection to be used. In order to achieve the desired results, the group are going to use the following tools: production of recombinant proteins and antibodies to these proteins, RNA interference, transcriptome, fluorescence microscopy, proteomic analysis, kinetic measurements, experiments in system of plasma resonance, immunization of cattle, experiments larvicides for Aedes and cell culture of tick and mosquito. The knowledge generated in this study will help in the understanding the role of protease inhibitors, proteases and other molecules in the vector-etiological agent interactions and also it will contribute to the formation of human resources and identification of molecules with biotechnological potential regarding problems related to livestock and public health. (AU)

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Scientific publications (17)
(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)
MANZATO, VERONICA MORAES; SOARES TORQUATO, RICARDO JOSE; ALVES LEMOS, FRANCISCO JOSE; NISHIDUKA, ERIKA; TASHIMA, ALEXANDRE KEIJI; TANAKA, APARECIDA SADAE. versatile inhibitor of digestive enzymes in Aedes aegypti larvae selected from a pacifastin (TiPI) phage display librar. Biochemical and Biophysical Research Communications, v. 590, p. 139-144, . (12/03657-8, 19/03779-5)
BUARQUE, DIEGO S.; GOMES, CICERA M.; ARAUJO, RICARDO N.; PEREIRA, MARCOS H.; FERREIRA, ROBERTA C.; GUARNERI, ALESSANDRA A.; TANAKA, APARECIDA S.. A new antimicrobial protein from the anterior midgut of Triatoma infestans mediates Trypanosoma cruzi establishment by controlling the microbiota. Biochimie, v. 123, p. 138-143, . (12/00723-0, 12/03657-8)
LU, STEPHEN; SOARES, TATIANE S.; VAZ JUNIOR, ITABAJARA S.; LOVATO, DIOGO V.; TANAKA, APARECIDA S.. Rmcystatin3, a cysteine protease inhibitor from Rhipicephalus microplus hemocytes involved in immune response. Biochimie, v. 106, p. 17-23, . (12/03657-8, 09/17589-1, 05/03514-9)
GOMES, CICERA MARIA; DE MORAIS-ZANI, KAREN; LU, STEPHEN; BUARQUE, DIEGO DE SOUZA; CARDOSO BRAZ, GLORIA REGINA; GREGO, KATHLEEN FERNANDES; TANAKA, APARECIDA SADAE; TANAKA-AZEVEDO, ANITA MITICO. Differential transcript profile of inhibitors with potential anti-venom role in the liver of juvenile and adult Bothrops jararaca snake. PeerJ, v. 5, . (14/11108-0, 12/03657-8, 13/05357-4)
SOARES, TATIANE S.; BUARQUE, DIEGO S.; QUEIROZ, BRUNA R.; GOMES, CICERA M.; BRAZ, GLORIA R. C.; ARAUJO, RICARDO N.; PEREIRA, MARCOS H.; GUARNERI, ALESSANDRA A.; TANAKA, APARECIDA S.. A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus. Biochimie, v. 112, p. 41-48, . (12/00723-0, 12/03657-8)
ABREU, PATRICIA A.; SOARES, TATIANE S.; BUARQUE, DIEGO S.; TORQUATO, RICARDO S.; TANAKA, APARECIDA S.. RmKK, a tissue kallikrein inhibitor from Rhipicephalus microplus eggs. Biochemical and Biophysical Research Communications, v. 449, n. 1, p. 69-73, . (12/03657-8, 05/03514-9)
LU, STEPHEN; DA ROCHA, LETICIA A.; TORQUATO, RICARDO J. S.; VAZ JUNIOR, ITABAJARA DA SILVA; FLORIN-CHRISTENSEN, MONICA; TANAKA, APARECIDA S.. A novel type 1 cystatin involved in the regulation of Rhipicephalus microplus midgut cysteine proteases. TICKS AND TICK-BORNE DISEASES, v. 11, n. 3, . (15/09268-1, 12/03657-8, 17/25609-9)
BUARQUE, DIEGO S.; BRAZ, GLORIA R. C.; MARTINS, RAFAEL M.; TANAKA-AZEVEDO, ANITA M.; GOMES, CICERA M.; OLIVEIRA, FELIPE A. A.; SCHENKMAN, SERGIO; TANAKA, APARECIDA S.. Differential Expression Profiles in the Midgut of Triatoma infestans Infected with Trypanosoma cruzi. PLoS One, v. 8, n. 5, . (12/03657-8, 12/00723-0, 05/03514-9)
LU, STEPHEN; ASCENCIO, MARIANO E.; TORQUATO, RICARDO J. S.; FLORIN-CHRISTENSEN, MONICA; TANAKA, APARECIDA S.. Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design. Biochimie, v. 179, p. 127-134, . (15/09268-1, 17/25609-9, 12/03657-8)
KARLA A. OLIVEIRA; RICARDO J. S. TORQUATO; DANIELA C. G. GARCIA LUSTOSA; TALES RIBEIRO; BRUNO W. L. NASCIMENTO; LILIAN C. G. DE OLIVEIRA; MARIA A. JULIANO; THAYSA PASCHOALIN; VIRGINIA S. LEMOS; RICARDO N. ARAUJO; et al. Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 27, . (12/03657-8)
SOARES, TATIANE S.; OLIVEIRA, FELIPE; TORQUATO, RICARDO J. S.; SASAKI, SERGIO D.; ARAUJO, MARIANA S.; PASCHOALIN, THAYSA; TANAKA, APARECIDA S.. BmTI-A, a Kunitz type inhibitor from Rhipicephalus microplus able to interfere in vessel formation. Veterinary Parasitology, v. 219, p. 44-52, . (12/03657-8, 05/03514-9)
TORQUATO, RICARDO J. S.; LU, STEPHEN; MARTINS, NADIA HELENA; TANAKA, APARECIDA S.; BARBOSA PEREIRA, PEDRO JOSE. High-resolution structure of a Kazal-type serine protease inhibitor from the dengue vector Aedes aegypti. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 73, n. 8, p. 469-475, . (12/03657-8)
SOARES, TATIANE SANCHES; RODRIGUEZ GONZALEZ, BORIS LUIS; SOARES TORQUATO, RICARDO JOSE; ALVES LEMOS, FRANCISCO JOSE; COSTA-DA-SILVA, ANDRE L.; CAPURRO GUIMARAES, MARGARETH DE LARA; TANAKA, APARECIDA SADAE. Functional characterization of a serine protease inhibitor modulated in the infection of the Aedes aegypti with dengue virus. Biochimie, v. 144, p. 160-168, . (12/15191-3, 12/03657-8)
SOARES, TATIANE SANCHES; SOARES TORQUATO, RICARDO JOSE; GONZALEZ, YAMILE GONZALEZ; ALVES LEMOS, FRANCISCO JOSE; TANAKA, APARECIDA SADAE. Production of serine protease inhibitors by mutagenesis and their effects on the mortality of Aedes aegypti L. larvae. PARASITES & VECTORS, v. 8, . (09/05405-3, 09/06003-6, 12/03657-8, 05/03514-9)
CARDOSO, THYAGO H. S.; LU, STEPHEN; GONZALEZ, BORIS R. G.; TORQUATO, RICARDO J. S.; TANAKA, APARECIDA S.. Characterization of a novel cystatin type 2 from Rhipicephalus microplus midgut. Biochimie, v. 140, p. 117-121, . (11/09340-3, 12/03657-8)
LU, S.; PARIZI, L. F.; TORQUATO, R. J. S.; VAZ JUNIOR, I. S.; TANAKA, A. S.. Novel pseudo-aspartic peptidase from the midgut of the tick Rhipicephalus microplus. SCIENTIFIC REPORTS, v. 9, . (15/09268-1, 12/03657-8)
OLIVEIRA, FELIPE A. A.; BURI, MARCUS V.; RODRIGUEZ, BORIS L.; COSTA-DA-SILVA, ANDRE L.; ARAUJO, HELENA R. C.; CAPURRO, MARGARETH L.; LU, STEPHEN; TANAKA, APARECIDA S.. The first characterization of a cystatin and a cathepsin L-like peptidase from Aedes aegypti and their possible role in DENV infection by the modulation of apoptosis. International Journal of Biological Macromolecules, v. 146, p. 141-149, . (13/16609-4, 12/03657-8)

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