Animal toxins: structure, function and biotechnological applications

• Grant number: 05/54855-0
• Support Opportunities: Research Projects - Thematic Grants
• Start date: March 01, 2007
• End date: May 31, 2011
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Suely Vilela
• Grantee: Suely Vilela
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Pesquisadores principais: Andreimar Martins Soares ; Eliane Candiani Arantes Braga ; Wagner Ferreira dos Santos
• Associated scholarship(s): 09/09829-2 - Assessment of cellular activation and inflammation induced by crude venom of the scorpion Tityus serrulatus and its fractions Ts1, Ts2, and Ts5 alone., BP.MS; 09/13902-7 - Characterization of synthetic and natural inhibitors of the human secreted phospholipase A2 of the group IIA, BP.PD; 09/14421-2 - Purification and structural and electrophysiological characterization of neurotoxins present in fraction VII-VIII from Tityus serrulatus venom, BP.IC; + associated scholarships 08/10760-4 - Functional and structural characterization of alfa-type phospholipase A2 inhibitor from B. alternatus snake plasma: cloning, expression and mapping of the region responsible for its inhibitory activity, BP.DR; 08/07373-9 - Structural and functional characterization of the precursor Ts1, the major toxin from Tityus serrulatus venom, BP.IC; 07/01021-0 - Functional and structural characterization of an acidic phospholipase A2 from Bothrops moojeni snake venom, BP.DD; 07/07710-2 - Snake Venom Phospholipases A2 Effect on Inflammation, BP.PD; 08/00977-6 - Structural Comparative Analysis of Toxic and Non-Toxic Acidic Phospholipase A2 Isolated from Snake Venoms., BP.PD; 07/08361-1 - Functional and structural characterization of a non-hemorrhagic metalloprotease with antitumor and antimicrobial actvity isolated from Bothrops pirajai snake venom, BP.DD; 07/06742-8 - Activity of neurotoxins isolated from the venom of Bufo paracnemis on PTZ-induced seziures and voltage-dependent Na+, K+ e Ca++ ion channels., BP.PD; 06/07177-0 - Evaluation of the potential antidengue and anti yellow fewer activity of animal toxins, BP.DR; 07/03626-7 - EVALUATION OF THE POTENTIAL ANTIDENGUE ACTIVITY OF ANIMAL TOXINS, BP.IC; 07/02726-8 - Structural and Functional Characterization of a non-complexed PLA2, inter-cro, Isolated from Crotalus durissus terrificus snake venom: Evaluation of Analgesic and antitumoral activities, BP.MS; 07/04741-4 - Study of activity of soluble hyaluronidases or encapsulated in association or not with MK886, BP.PD - associated scholarships

Abstract

Researches on animal toxins have contributed significantly with the development of Biological Sciences. An extensive study involving evaluation of the effects of venoms and their toxins upon different biological systems, relating them with the structure of active components, will be able to allow the development of new experimental tools and for important therapeutic agents inc1uding neuroprotectors, neurotoxins, immunomodulators and c10tting or anticoagulant factors. These studies will also be able to help in the elucidation of the c1inical picture produced by envenoming, opening the way for a more effective therapeutic strategy. hemostasis unbalance. Gene Cox-2, strictly involved with the inflammatory process, codes for the cyc10xigenase enzyme which catalyzes the transformation of arachidonic acid to prostaglandins. On basis of these considerations, our project is aimed at the investigation of the behavior of the PC 12 cell line regarding its cell cyc1e, apoptosis and expression of genes involved with signaling of injuries and inflammatory process, by means of PCR at real time, resulting from treatment with different toxins. Analysis of the activation pathways of COX-2 through study of gene expression will lead to a better understanding of this path in response to treatment with these toxins. In addition, the proteome analysis of these cells will be carried out in order to detect alterations in the protein synthesis induced by the toxins. Isolation of the biologically active components will be performed with use of c1assical chromatographic procedures, namely gel filtration, ionic exchange, hydrophobic interaction, bioaffinity and CLAE (reverse phase). This stage of the project is fundamental for all other subsequent stages, since it will provide the active components which will be, together with the crude venom, the purpose of study of this work. Enzymatic activities (hyaluronidase, proteolytic, PLA2, nuc1eotidase, L-aminoacid oxidase), biological (hemorrhagic, myotoxic, edema inducing, coagulant, anticoagulant, platelet aggregating), immunomodulatory (effect upon the complement system, anti-inflammatory, cell activation marking, apoptoic potential of the venom components), microbicide and cytotoxic (leishmanicide, trypanosomicidal, bactericidal, antitumoral, antiviral and fungicidal) and lectin activity of venoms or their isolated components will be studied. All these activities were chosen because they are venom targets and highly relevant in physiological processes. Within the different systems to be evaluated, the structure of isolated toxins will be the basis for elucidation of the structure-function relationship. The structural characterization of the toxins and their interactions with ligands and receptors will be performed through direct sequencing of the proteins, together with the cloning and sequencing of their corresponding DNA, X-ray crystallography and molecular modeling. The yet not elucidated structure of a protein can be predicted by modeling and homology, using known structures of other proteins with which it relates along evolution through a common ancestral. In the last years, modeling by molecular homology was shown to be highly important in Medicinal Chemistry, specifically plaining the development of new drugs. Keeping in mind the number of venoms and their active components to be studied, as well as the proposed assays for the characterization of their effects and structures, we have to consider this work as multidisciplinary, extensive and promising. Finally, results of this project certainly will broad the knowledge on structure-function relationship of toxins in biological systems, with probable applications in medical clinics and basic research. (AU)

Scientific publications

(13)

(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)

BERNARDES, CAROLINA P.; MENALDO, DANILO L.; MAMEDE, CARLA C. N.; ZOCCAL, KARINA F.; CINTRA, ADELIA C. O.; FACCIOLI, LUCIA H.; STANZIOLA, LEONILDA; DE OLIVEIRA, FABIO; SAMPAIO, SUELY V.. Evaluation of the local inflammatory events induced by BpirMP, a metalloproteinase from Bothrops pirajai venom. Molecular Immunology, v. 68, n. 2, B, p. 456-464, 2015. (07/08361-1, 05/54855-0, 11/23236-4)

ALVES, RAQUEL MELO; FELICIANO, PATRICIA ROSA; SAMPAIO, SUELY VILELA; NONATO, MARIA CRISTINA. A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 67, n. 4, p. 475-478, APR 2011. (05/54855-0, 09/10454-3)

SANTUSSI, WELLIGTON M.; BORDON, KARLA C. F.; RODRIGUES ALVES, ANA P. N.; COLOGNA, CAMILA T.; SAID, SURAIA; ARANTES, ELIANE C.. Antifungal Activity against Filamentous Fungi of Ts1, a Multifunctional Toxin from Tityus serrulatus Scorpion Venom. FRONTIERS IN MICROBIOLOGY, v. 8, JUN 6 2017. (15/18432-0, 05/54855-0)

MULLER, VANESSA DANIELLE; SOARES, RICARDO OLIVEIRA; DOS SANTOS-JUNIOR, NILTON NASCIMENTO; TRABUCO, AMANDA CRISTINA; CINTRA, ADELIA CRISTINA; FIGUEIREDO, LUIZ TADEU; CALIRI, ANTONIO; SAMPAIO, SUELY VILELA; AQUINO, VICTOR HUGO. Phospholipase A(2) Isolated from the Venom of Crotalus durissus terrificus Inactivates Dengue virus and Other Enveloped Viruses by Disrupting the Viral Envelope. PLoS One, v. 9, n. 11, NOV 10 2014. (08/50617-6, 05/54855-0)

BREGGE-SILVA, CRISTIANE; NONATO, MARIA CRISTINA; DE ALBUQUERQUE, SERGIO; HO, PAULO LEE; JUNQUEIRA DE AZEVEDO, INACIO L. M.; VASCONCELOS DINIZ, MARCELO RIBEIRO; LOMONTE, BRUNO; RUCAVADO, ALEXANDRA; DIAZ, CECILIA; MARIA GUTIERREZ, JOSE; et al. Isolation and biochemical, functional and structural characterization of a novel L-amino acid oxidase from Lachesis muta snake venom. Toxicon, v. 60, n. 7, p. 1263-1276, DEC 1 2012. (05/54855-0)

MENDONCA-FRANQUEIRO, ELAINE DE PAULA; ALVES-PAIVA, RAQUEL DE MELO; SARTIM, MARCO AURELIO; CALLEJON, DANIEL ROBERTO; PAIVA, HELDER HENRIQUE; ANTONUCCI, GILMARA AUSECH; ROSA, JOSE CESAR; OLIVEIRA CINTRA, ADELIA CRISTINA; FRANCO, JOAO JOSE; ARANTES, ELIANE CANDIANI; et al. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom. ACTA BIOCHIMICA ET BIOPHYSICA SINICA, v. 43, n. 3, p. 181-192, 2011. (05/54855-0)

BERNARDES, CAROLINA P.; MENALDO, DANILO L.; MAMEDE, CARLA C. N.; ZOCCAL, KARINA F.; CINTRA, ADELIA C. O.; FACCIOLI, LUCIA H.; STANZIOLA, LEONILDA; DE OLIVEIRA, FABIO; SAMPAIO, SUELY V.. Evaluation of the local inflammatory events induced by BpirMP, a metalloproteinase from Bothrops pirajai venom. Molecular Immunology, v. 68, n. 2, p. 9-pg., 2015-01-01. (11/23236-4, 07/08361-1, 05/54855-0)

COLOGNA, CAMILA T.; PEIGNEUR, STEVE; RUSTIGUEL, JOANE K.; NONATO, M. CRISTINA; TYTGAT, JAN; ARANTES, ELIANE C.. Investigation of the relationship between the structure and function of Ts2, a neurotoxin from Tityus serrulatus venom. FEBS Journal, v. 279, n. 8, p. 1495-1504, APR 2012. (05/54855-0)

DE TONI, LANUZE G. B.; MENALDO, DANILO L.; CINTRA, ADELIA C. O.; FIGUEIREDO, MARIA J.; DE SOUZA, ANDERSON R.; MAXIMIANO, WILLIAM M. A.; JAMUR, MARIA C.; SOUZA, GLORIA E. P.; SAMPAIO, SUELY V.. Inflammatory mediators involved in the paw edema and hyperalgesia induced by Batroxase, a metalloproteinase isolated from Bothrops atrox snake venom. International Immunopharmacology, v. 28, n. 1, p. 199-207, SEP 2015. (05/54855-0)

ZOCCAL, KARINA FURLANI; BITENCOURT, CLAUDIA DA SILVA; SECATTO, ADRIANA; SORGI, CARLOS ARTERIO; FIGUEREDO BORDON, KARLA DE CASTRO; SAMPAIO, SUELY VILELA; ARANTES, ELIANE CANDIANI; FACCIOLI, LUCIA HELENA. Tityus serrulatus venom and toxins Ts1, Ts2 and Ts6 induce macrophage activation and production of immune mediators. Toxicon, v. 57, n. 7-8, p. 1101-1108, JUN 2011. (05/54855-0, 09/07169-5, 09/09829-2)

BITENCOURT, CLAUDIA DA SILVA; DA SILVA, LETICIA BUENO; TARTARI PEREIRA, PRISCILLA APARECIDA; GELFUSO, GUILHERME MARTINS; FACCIOLI, LUCIA HELENA. Microspheres prepared with different co-polymers of poly(lactic-glycolic acid) (PLGA) or with chitosan cause distinct effects on macrophages. COLLOIDS AND SURFACES B-BIOINTERFACES, v. 136, p. 678-686, DEC 1 2015. (07/04741-4, 02/12856-2, 09/07169-5, 05/54855-0)

ALVES, RAQUEL MELO; FELICIANO, PATRICIA ROSA; SAMPAIO, SUELY VILELA; NONATO, MARIA CRISTINA. A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 67, p. 4-pg., 2011-04-01. (05/54855-0, 09/10454-3)

CREMONEZ, CAROLINE MARRONI; LEITE, FLAVIA PINE; FIGUEIREDO BORDON, KARLA DE CASTRO; CERNI, FELIPE AUGUSTO; CARDOSO, IARA AIME; DE OLIVEIRA GREGORIO, ZITA MARIA; GONCALVES DE SOUZA, RODRIGO CANCADO; DE SOUZA, ANA MARIA; ARANTES, ELIANE CANDIANI. Experimental Lachesis muta rhombeata envenomation and effects of soursop (Annona muricata) as natural antivenom. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 22, MAR 8 2016. (05/54855-0, 12/13590-8)

Filed patent(s) as a result of this research project