New functional perspectives of a serine protease from Crotalus durissus collilineatus: activity on voltage-gated ion channels

• Grant number: 14/16182-3
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): October 01, 2014
• Effective date (End): March 31, 2018
• Field of knowledge: Health Sciences - Pharmacy - Toxicological Analysis
• Principal researcher: Eliane Candiani Arantes Braga
• Grantee: Johara Boldrini França
• Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis, AP.TEM
• Associated scholarship(s): 15/16714-8 - New functional perspectives of a serine protease from Crotalus durissus collilineatus: activity on voltage-gated ion channels, BE.EP.PD

Abstract

The complexity and variety of physiological targets with which snake venom toxins interact make them important candidates for models in the development of new drugs and biological tools. The venoms from Viperidae snakes are rich sources of proteolytic enzymes that exert various biological activities in the prey or victim, such as activation of the complement system and inflammation, changes in cell differentiation, interactions with the hemostatic system, among others. Snake venom serine proteases (SVSPs) act mainly on plasma proteins, generating a variety of physiological changes with effects on platelet aggregation, blood coagulation, fibrinolysis, blood pressure, complement system and nervous system. However, the complexity of the mechanisms by which SVSPs affect the homeostasis of the victim was not yet completely clarified. Therefore, this project aims to find new perspectives on the functionality of a SVSP from Crotalus durissus collilineatus, called collinein-1, in order to expand the fields of application of this class of toxins. In this context, we propose to investigate the action of collinein-1 in native, recombinant and mutant forms on different voltage-gated ion channels. Native collinein-1 will be purified from venom by a previously described chromatographic method and the recombinant enzyme will be expressed in Pichia pastoris in catalytically active and inactive forms (presenting a replacement of His43 by Arg43), according to previously standardized protocol. Toxins will be tested for their effect on various voltage-gated ion channels (Kv and Nav) expressed in Xenopus laevis oocytes by electrophysiology assays. The mechanisms by which collinein-1 acts on the voltage-gated ion channels will be investigated by electrophysiological assays using catalytically inactive toxin and by testing the reversibility of collinein-1 blocking effect. Ionic channels play important cellular functions such as excitation of nerves and muscle cells, regulation of blood pressure, mobility and sperm capacitation and modulation of tumor cell proliferation. Despite the importance of these channels, there are few snake toxins that was already studied for their action on ion channels, which reinforces the importance of the proposed study in the search for new ion channel modulating or blocking toxins with potential application in various areas of science and/or health.

Scientific publications

(4)

(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)

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. (16/04761-4, 15/17286-0, 11/23236-4, 17/13485-3, 17/14035-1, 15/00740-0, 18/14158-9, 15/18432-0, 16/24191-8, 14/16182-3, 15/16714-8)

AMORIM, FERNANDA GOBBI; CORDEIRO, FRANCIELLE ALMEIDA; PINHEIRO-JUNIOR, ERNESTO LOPES; BOLDRINI-FRANCA, JOHARA; ARANTES, ELIANE CANDIANI. Microbial production of toxins from the scorpion venom: properties and applications. Applied Microbiology and Biotechnology, v. 102, n. 15, p. 6319-6331, AUG 2018. (14/16182-3, 16/04761-4, 12/14996-8, 11/12317-3)

PINHEIRO-JUNIOR, ERNESTO LOPES; BOLDRINI-FRANCA, JOHARA; PIRES DE CAMPOS ARAUJO, LUCIANA MATTOSO; SANTOS-FILHO, NORIVAL ALVES; BENDHACK, LUSIANE MARIA; CILLI, EDUARDO MAFFUD; ARANTES, ELIANE CANDIANI. LmrBPP9: A synthetic bradykinin-potentiating peptide from Lachesis muta rhombeata venom that inhibits the angiotensin-converting enzyme activity in vitro and reduces the blood pressure of hypertensive rats. Peptides, v. 102, p. 1-7, APR 2018. (14/05538-1, 14/16182-3, 13/07600-3)

DE OLIVEIRA, ISADORA SOUSA; CARDOSO, IARA AIME; FIGUEIREDO BORDON, KARLA DE CASTRO; IMAI CARONE, SANTE EMMANUEL; BOLDRINI-FRANCA, JOHARA; PUCCA, MANUELA BERTO; ZOCCAL, KARMA FURLANI; FACCIOLI, LUCIA HELENA; SAMPAIO, SUELY VILELA; ROSA, JOSE CESAR; et al. Global proteomic and functional analysis of Crotalus durissus collilineatus individual venom variation and its impact on envenoming. JOURNAL OF PROTEOMICS, v. 191, n. SI, p. 153-165, JAN 16 2019. (14/16182-3, 14/03332-7, 11/23236-4)