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Studies of the physiological function and biotechnological potential of protease inhibitors and anti-hemostatics in hematophagous arthropods


The biological molecules diversity present in hematophagous arthropods, in addition to their importance as disease vectors have motivated our group to investigate new anti-hemostatic and protease inhibitors present in three different arthropod species which are responsible for disease transmission to humans and animals in Brazil. They are: Triatoma infestans kissing bug, Boophilus microplus bovine tick and Aedes aegypti mosquito. In these three cases, the uncontrolled use of pesticide to control those vectors has caused the environment and human contaminations. Long term pesticide exposition of ectoparasite has resulted to resistance problems. The World Health Organization has suggested that the main cause of unsuccess in these disease controls is our ignorance in the biology of these animals. Therefore, a better comprehension of several mechanisms which allowed these ectoparasites successful adaptation for hematophagy would reveal new targets to control strategy, vaccine and also new drugs developments. During the last eight years, our group has been contributing for better understanding of some hematophagous arthropods biology, among them the T. infestans bug which control its host blood coagulation by thrombin and factor XIIa inhibitors, and interestingly they are coding by the same cDNA precursor molecule. Our studies using factor XIIa inhibitor allowed a patent process submission. We also showed several serine protease inhibitors from B. microplus tick and H. irritans fly which could present physiological function such as controlling endogenous proteases and or microorganisms. In the present project our aims will be the functional and structural characterization of several molecules identified by our group, to reach these objectives we are going to use the following tools: RNA interference, PCR real time, proteome, antifungal experiments, microorganism protease inhibitions, hemolymph coagulation and phenoloxidase cascade activations. With these efforts we intend to understand these molecules role in the hematophagous physiology and also use these molecules: for arthropod protein-protein interaction understanding, for vaccine development and biotechnology potential molecule to be used in human and veterinary medicines and agriculture. (AU)

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Scientific publications (20)
(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)
WATANABE, RENATA M. O.; SOARES, TATIANE S.; MORAIS-ZANI, KAREN; TANAKA-AZEVEDO, ANITA M.; MACIEL, CERES; CAPURRO, MARGARETH L.; TORQUATO, RICARDO J. S.; TANAKA, APARECIDA S.. A novel trypsin Kazal-type inhibitor from Aedes aegypti with thrombin coagulant inhibitory activity. Biochimie, v. 92, n. 8, p. 933-939, . (05/03514-9)
DE MARCO, RENATO; AZZOLINI, SIMONE S.; LOVATO, DIOGO V.; TORQUATO, RICARDO J. S.; AMINO, ROGERIO; DE MIRANDA, ANTONIO; TANAKA, APARECIDA S.. Validation of a Phage Display Method for Protease Inhibitor Selection Using SFTI and HiTI Synthetic Hybrid Peptides. COMBINATORIAL CHEMISTRY & HIGH THROUGHPUT SCREENING, v. 13, n. 9, p. 829-835, . (05/03514-9)
SOARES‚ T.S.; WATANABE‚ R.M.O.; LEMOS‚ F.J.A.; TANAKA‚ A.S.. Molecular characterization of genes encoding trypsin-like enzymes from< i> Aedes aegypti larvae and identification of digestive enzymes. GENESIS, v. 489, n. 2, p. 70-75, . (05/03514-9)
SOARES, TATIANE SANCHES; SOARES TORQUATO, RICARDO JOSE; ALVES LEMOS, FRANCISCO JOSE; TANAKA, APARECIDA SADAE. Selective inhibitors of digestive enzymes from Aedes aegypti larvae identified by phage display. Insect Biochemistry and Molecular Biology, v. 43, n. 1, p. 9-16, . (09/05405-3, 05/03514-9)
SOARES, TATIANE S.; WATANABE, RENATA M. O.; LEMOS, FRANCISCO J. A.; TANAKA, APARECIDA S.. Molecular characterization of genes encoding trypsin-like enzymes from Aedes aegypti larvae and identification of digestive enzymes. Gene, v. 489, n. 2, p. 70-75, . (05/03514-9)
CAMPOS, I. T. N.; SOUZA, T. A. C. B.; TORQUATO, R. J. S.; DE MARCO, R.; TANAKA-AZEVEDO, A. M.; TANAKA, A. S.; BARBOSA, J. A. R. G.. The Kazal-type inhibitors infestins 1 and 4 differ in specificity but are similar in three-dimensional structure. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, v. 68, n. 6, p. 695-702, . (01/07535-0, 05/03514-9)
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)
WATANABE, RENATA M. O.; TANAKA-AZEVEDO, ANITA M.; ARAUJO, MARIANA S.; JULIANO, MARIA A.; TANAKA, APARECIDA S.. Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity. Biochimie, v. 93, n. 3, p. 618-623, . (05/03514-9)
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)
BUARQUE, DIEGO S.; SPINDOLA, LETICIA M. N.; MARTINS, RAFAEL M.; BRAZ, GLORIA R. C.; TANAKA, APARECIDA S.. Tigutcystatin, a cysteine protease inhibitor from Triatoma infestans midgut expressed in response to Trypanosoma cruzi. Biochemical and Biophysical Research Communications, v. 413, n. 2, p. 241-247, . (05/03514-9)
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)
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)
SASAKI‚ S. D.; TANAKA‚ A. S.. rBmTI-6‚ a Kunitz-BPTI domain protease inhibitor from the tick Boophilus microplus, its cloning‚ expression and biochemical characterization. Veterinary Parasitology, v. 155, n. 1/2, p. 133-141, . (05/03514-9)
LIMA, CASSIA A.; TORQUATO, RICARDO J. S.; SASAKI, SERGIO D.; JUSTO, GISELLE Z.; TANAKA, APARECIDA S.. Biochemical characterization of a Kunitz type inhibitor similar to dendrotoxins produced by Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) hemocytes. Veterinary Parasitology, v. 167, n. 2-4, SI, p. 279-287, . (05/03514-9)
CLARA, RENAN O.; SOARES, TATIANE S.; TORQUATO, RICARDO J. S.; LIMA, CASSIA A.; WATANABE, RENATA O. M.; BARROS, NILANA M. T.; CARMONA, ADRIANA K.; MASUDA, AOI; VAZ JUNIOR, ITABAJARA S.; TANAKA, APARECIDA S.. Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: Specificity study using a peptide phage display library. Veterinary Parasitology, v. 181, n. 2-4, p. 291-300, . (05/03514-9)
SOARES, TATIANE SANCHES; OKUTA WATANABE, RENATA MIDORI; TANAKA-AZEVEDO, ANITA MITICO; SOARES TORQUATO, RICARDO JOSE; LU, STEPHEN; FIGUEIREDO, ANA CARVALHO; BARBOSA PEREIRA, PEDRO JOSE; TANAKA, APARECIDA S.. Expression and functional characterization of boophilin, a thrombin inhibitor from Rhipicephalus (Boophilus) microplus midgut. Veterinary Parasitology, v. 187, n. 3-4, p. 521-528, . (09/05405-3, 05/03514-9)
DE MARCO, RENATO; LOVATO, DIOGO V.; TORQUATO, RICARDO J. S.; CLARA, RENAN O.; BUARQUE, DIEGO S.; TANAKA, APARECIDA S.. The first pacifastin elastase inhibitor characterized from a blood sucking animal. Peptides, v. 31, n. 7, p. 1280-1286, . (05/03514-9)
SASAKI, SERGIO D.; LIMA, CÁSSIA A. DE; LOVATO, DIOGO V.; JULIANO, MARIA A.; TORQUATO, RICARDO J. S.; TANAKA, APARECIDA S.. BmSI-7, a novel subtilisin inhibitor from Boophilus microplus, with activity toward Pr1 proteases from the fungus Metarhizium anisopliae. Experimental Parasitology, v. 118, n. 2, p. 214-220, . (05/03514-9)
LOVATO, DIOGO V.; AMINO, ROGERIO; GONZALEZ, YAMILE; MIRANDA, ANTONIO; SCHENKMAN, SERGIO; TANAKA, APARECIDA S.. Infestin 1R, an intestinal subtilisin inhibitor from Triatoma infestans able to impair mammalian cell invasion by Trypanosoma cruzi. Experimental Parasitology, v. 129, n. 4, p. 362-367, . (05/03514-9)
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)

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