Structural and functional studies aiming to understand the role of snake venoms toxins and how to inhibit their biological activity

• Grant number: 15/17286-0
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2016
• End date: March 31, 2019
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Marcos Roberto de Mattos Fontes
• Grantee: Marcos Roberto de Mattos Fontes
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated researchers: Angelo José Magro ; Maeli Dal Pai ; Marcia Gallacci ; Ney Lemke ; Paulo Correia-de-Sá

Abstract

Envenoming caused by snakebites is considered a public health problem in many tropical and subtropical countries and has recently been included in the list of Neglected Tropical Diseases by the World Health Organization. In Latin America, the majority of ophidic accidents are caused by snakes from Bothrops and Crotalus genus. One of the main complications regarding bothropic envenomation is the prominent local muscle damage and subsequent neuromuscular transmission block, which is usually induced by phospholipase A2, an important component of these venoms. In this work, we propose for the first time, a broad structural and functional study of snake venom proteins - native, recombinant and complexed to potential inhibitors. The following techniques will be used: i) structural/biophysical: crystallography, small angle X-ray scattering, circular dichroism, dynamic light scattering, fluorescence and isothermal titration calorimetry, structural bioinformatics; ii) in vitro functional studies including electrophysiology, myographic techniques, real-time video-microscopy and radiochemical studies; iii) morphologic analysis (optic, confocal and electronic microscopy). This information could be very important for the identification, characterization and development (drug design) of substances with biotechnological and pharmacological usefulness. These compounds may be used for the control of symptoms that cannot be prevented or reverted by serum therapy administration and as aid for the treatment of the underlying pathological process. This project is aligned with other projects previously supported by FAPESP (four grants in the snake venom proteins field) and CNPq (three grants in the field) since 1997. Furthermore, the Brazilian laboratory was also supported by INCT in Toxinology (FAPESP/CNPq) and Toxinology project of the CAPES agency. The scientific output of the Brazilian laboratory in the snake venom toxin field accounts for more than 60 papers and 16 concluded theses in this field. Since 1991, the Laboratory do Pharmacology and Neurobiology, ICBAS-UPorto has done pioneering research in the study of the pathogenesis of neuromuscular transmission deficits, routinely using neurochemical, electrophysiological and bioimaging approaches such as those required for this project. Both Brazilian and Portuguese research groups have a fruitful collaborative project which includes a recently published paper within the frame of a post-doctoral grant by FAPESP (BEPE-FAPESP n° 2013/03624-5). (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)

SALVADOR, GUILHERME H. M.; DOS SANTOS, JULIANA I.; BORGES, RAFAEL J.; FONTES, MARCOS R. M.. Structural evidence for a fatty acid-independent myotoxic mechanism for a phospholipase A(2)-like toxin. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1866, n. 3, p. 473-481, MAR 2018. (15/17286-0)

BOLDRINI-FRANCA, JOHARA; PINHEIRO-JUNIOR, ERNESTO LOPES; PEIGNEUR, STEVE; PUCCA, MANUELA BERTO; CERNI, FELIPE AUGUSTO; BORGES, RAFAEL JUNQUEIRA; COSTA, TASSIA RAFAELLA; IMAI CARONE, SANTE EMMANUEL; DE MATTOS FONTES, MARCOS ROBERTO; SAMPAIO, SUELY VILELA; et al. Beyond hemostasis: a snake venom serine protease with potassium channel blocking and potential antitumor activities. SCIENTIFIC REPORTS, v. 10, n. 1, MAR 11 2020. (15/18432-0, 15/00740-0, 18/14158-9, 16/24191-8, 15/16714-8, 17/13485-3, 11/23236-4, 17/14035-1, 16/04761-4, 14/16182-3, 15/17286-0)

CARDOSO, FABIO F.; DE SOUZA, MAXIMILIA F.; OLIVEIRA, CRISTIANO L. P.; FONTES, MARCOS R. M.. In-solution structural studies involving a phospholipase A 2-like myotoxin and a natural inhibitor: Plasticity of oligomeric assembly affects mechanisms of inhibition. Biochimie, v. 181, p. 145-153, FEB 2021. (15/17286-0, 12/07112-6, 18/16092-5)

SALVADOR, GUILHERME H. M.; BORGES, RAFAEL J.; LOMONTE, BRUNO; LEWIN, MATTHEW R.; FONTES, MARCOS R. M.. The synthetic varespladib molecule is a multi-functional inhibitor for PLA(2) and PLA(2)-like ophidic toxins. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1865, n. 7, JUL 2021. (15/17286-0, 16/24191-8)

BORGES, RAFAEL J.; CARDOSO, FABIO F.; FERNANDES, CARLOS A. H.; DREYER, THIAGO R.; DE MORAES, DELKIA S.; FLORIANO, RAFAEL S.; RODRIGUES-SIMIONI, LEA; FONTES, MARCOS R. M.. Functional and structural studies of a Phospholipase A(2)-like protein complexed to zinc ions: Insights on its myotoxicity and inhibition mechanism. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1861, n. 1, p. 11-pg., 2017-01-01. (15/17286-0, 13/00873-4)

SALVADOR, GUILHERME H. M.; DOS SANTOS, JULIANA I.; LOMONTE, BRUNO; FONTES, MARCOS R. M.. Crystal structure of a phospholipase A(2) from Bothrops asper venom: Insights into a new putative "myotoxic cluster". Biochimie, v. 133, p. 8-pg., 2017-02-01. (15/17286-0, 15/24167-7)

BORGES, RAFAEL J.; CARDOSO, FABIO F.; FERNANDES, CARLOS A. H.; DREYER, THIAGO R.; DE MORAES, DELKIA S.; FLORIANO, RAFAEL S.; RODRIGUES-SIMIONI, LEA; FONTES, MARCOS R. M.. Functional and structural studies of a Phospholipase A(2)-like protein complexed to zinc ions: Insights on its myotoxicity and inhibition mechanism. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1861, n. 1, A, p. 3199-3209, JAN 2017. (13/00873-4, 15/17286-0)

SALVADOR, GUILHERME H. M.; DOS SANTOS, JULIANA I.; LOMONTE, BRUNO; FONTES, MARCOS R. M.. Crystal structure of a phospholipase A(2) from Bothrops asper venom: Insights into a new putative ``myotoxic cluster{''}. Biochimie, v. 133, p. 95-102, FEB 2017. (15/24167-7, 15/17286-0)

FERNANDES, CARLOS A. H.; PAZIN, WALLANCE M.; DREYER, THIAGO R.; BICEV, RENATA N.; CAVALCANTE, WALTER L. G.; FORTES-DIAS, CONSUELO L.; ITO, AMANDO S.; OLIVEIRA, CRISTIANO L. P.; FERNANDEZ, ROBERTO MORATO; FONTES, MARCOS R. M.. Biophysical studies suggest a new structural arrangement of crotoxin and provide insights into its toxic mechanism. SCIENTIFIC REPORTS, v. 7, MAR 3 2017. (15/17286-0, 13/17864-8)

CARDOSO, FABIO F.; GOMES, ANTONIEL A. S.; DREYER, THIAGO R.; CAVALCANTE, WALTER L. G.; DAL PAI, MAELI; GALLACCI, MARCIA; FONTES, MARCOS R. M.. Neutralization of a bothropic PLA(2)-like protein by caftaric acid, a novel potent inhibitor of ophidian myotoxicity. Biochimie, v. 170, p. 163-172, MAR 2020. (12/07112-6, 15/17286-0)

FERNANDES, CARLOS A. H.; MOREA, EDNA GICELA O.; DOS SANTOS, GABRIEL A.; DA SILVA, VITOR L.; VIEIRA, MARINA ROVERI; VIVIESCAS, MARIA ALEJANDRA; CHATAIN, JEAN; VADEL, AURELIE; SAINTOME, CAROLE; FONTES, MARCOS ROBERTO M.; et al. A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1864, n. 7, JUL 2020. (15/17286-0, 18/04375-2, 19/11496-3)

BORGES, RAFAEL J.; SALVADOR, GUILHERME H. M.; PIMENTA, DANIEL C.; DOS SANTOS, LUCILENE D.; FONTES, MARCOS R. M.; USON, ISABEL. SEQUENCE SLIDER: integration of structural and genetic data to characterize isoforms from natural sources. Nucleic Acids Research, v. 50, n. 9, p. 16-pg., 2022-05-20. (16/24191-8, 15/17286-0, 19/05958-4, 20/10143-7)

CARDOSO, FABIO F.; BORGES, RAFAEL J.; DREYER, THIAGO R.; SALVADOR, GUILHERME H. M.; CAVALCANTE, WALTER L. G.; DAL PAI, MAELI; GALLACCI, MARCIA; FONTES, MARCOS R. M.. Structural basis of phospholipase A(2)-like myotoxin inhibition by chicoric acid, a novel potent inhibitor of ophidian toxins. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1862, n. 12, p. 2728-2737, DEC 2018. (15/17286-0, 12/07112-6)