Beyond hemostasis: a snake venom serine protease with potassium channel blocking and potential antitumor activities

Author(s):Show less -

Boldrini-Franca, Johara ^{[1, 2]} ; Pinheiro-Junior, Ernesto Lopes ^{[1, 3]} ; Peigneur, Steve ^[3] ; Pucca, Manuela Berto ^[4] ; Cerni, Felipe Augusto ^[1] ; Borges, Rafael Junqueira ^[5] ; Costa, Tassia Rafaella ^[1] ; Imai Carone, Sante Emmanuel ^[1] ; de Mattos Fontes, Marcos Roberto ^[5] ; Sampaio, Suely Vilela ^[1] ; Arantes, Eliane Candiani ^[1] ; Tytgat, Jan ^[3]

Affiliation

^[1] Univ Sao Paulo, Sch Pharmaceut Sci Ribeirao Preto, Av Cafe S-N, BR-14040903 Ribeirao Preto, SP - Brazil

^[2] Univ Vila Velha, Av Comissario Jose Dantas de Melo 21, Boa Vista 2, BR-29102920 Vila Velha, ES - Brazil

^[3] Katholieke Univ Leuven, Toxicol & Pharmacol, O&N 2 Herestr 49, B-3000 Leuven - Belgium

^[4] Univ Fed Roraima, Med Sch Roraima, Av Capitao Ene Garcez 2413, BR-69310970 Boa Vista, Parana - Brazil

^[5] Sao Paulo State Univ, UNESP, Inst Biosci, Rua Prof Dr Antonio Celso Wagner Zanin250, BR-18618689 Botucatu, SP - Brazil

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 10, n. 1 MAR 11 2020.
• Web of Science Citations: 1

Abstract

Snake venom serine proteases (SVSPs) are complex and multifunctional enzymes, acting primarily on hemostasis. In this work, we report the hitherto unknown inhibitory effect of a SVSP, named collinein-1, isolated from the venom of Crotalus durissus collilineatus, on a cancer-relevant voltage-gated potassium channel (hEAG1). Among 12 voltage-gated ion channels tested, collinein-1 selectively inhibited hEAG1 currents, with a mechanism independent of its enzymatic activity. Corroboratively, we demonstrated that collinein-1 reduced the viability of human breast cancer cell line MCF7 (high expression of hEAG1), but does not affect the liver carcinoma and the non-tumorigenic epithelial breast cell lines (HepG2 and MCF10A, respectively), which present low expression of hEAG1. In order to obtain both functional and structural validation of this unexpected discovery, where an unusually large ligand acts as an inhibitor of an ion channel, a recombinant and catalytically inactive mutant of collinein-1 (His43Arg) was produced and found to preserve its capability to inhibit hEAG1. A molecular docking model was proposed in which Arg79 of the SVSP 99-loop interacts directly with the potassium selectivity filter of the hEAG1 channel. (AU)

• FAPESP's process: 15/18432-0 - Bioprospection of animal toxins with biotechnological interest through omic tools
• Grantee: Eliane Candiani Arantes Braga
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/00740-0 - Therapeutic potential evaluation of L-amino acid oxidases isolated from snake venoms as antitumor: genotoxicity and gene expression studies
• Grantee: Tássia Rafaella Costa
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 18/14158-9 - Improving of human fragment antibodies (scFvs) specific for animals' venoms
• Grantee: Felipe Augusto Cerni
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 16/24191-8 - Development of methods for crystallographic structure elucidation and structural studies of toxic mechanism of snake venom Phospholipases A2 homologue proteins
• Grantee: Rafael Junqueira Borges
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 15/16714-8 - New functional perspectives of a serine protease from Crotalus durissus collilineatus: activity on voltage-gated ion channels
• Grantee: Johara Boldrini França Stringari
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 17/13485-3 - Development and release of SEQUENCE SLIDER: a multi-side chain evaluator applied to toxinology and phasing
• Grantee: Rafael Junqueira Borges
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis
• Grantee: Suely Vilela
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/14035-1 - Improving of human fragment antibodies (scFvs) specific for animals' venoms
• Grantee: Felipe Augusto Cerni
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 16/04761-4 - Functional and structural characterization and immune response evaluation of a recombinant serine protease from Crotalus durissus collilineatus modified by PEGylation
• Grantee: Ernesto Lopes Pinheiro Junior
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 14/16182-3 - New functional perspectives of a serine protease from Crotalus durissus collilineatus: activity on voltage-gated ion channels
• Grantee: Johara Boldrini França Stringari
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 15/17286-0 - Structural and functional studies aiming to understand the role of snake venoms toxins and how to inhibit their biological activity
• Grantee: Marcos Roberto de Mattos Fontes
• Support Opportunities: Regular Research Grants