Proteome of fraction from Tityus serrulatus venom reveals new enzymes and toxins

Author(s)

Amorim, Fernanda Gobbi ^{[1, 2]} ; Longhim, Heloisa Tavoni ^[2] ; Cologna, Camila Takeno ^{[2, 3]} ; Degueldre, Michel ^[3] ; De Pauw, Edwin ^[3] ; Quinton, Loic ^[3] ; Arantes, Eliane Candiani ^[2]

Affiliation

^[1] Univ Vila Velha, Vila Velha, ES - Brazil

^[2] Univ Sao Paulo, Dept Phys & Chem, Sch Pharmaceut Sci Ribeirao Preto, Av Cafe S-N, BR-14040903 Ribeirao Preto, SP - Brazil

^[3] Univ Liege, MolSys Res Unit, Lab Mass Spectrometry, Liege - Belgium

• Document type: Journal article
• Source: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 25, APR 18 2019.
• Web of Science Citations: 1

Abstract

Background: Tityus serrulatus venom (Ts venom) is a complex mixture of several compounds with biotechnological and therapeutical potentials, which highlights the importance of the identification and characterization of these components. Although a considerable number of studies have been dedicated to the characterization of this complex cocktail, there is still a limitation of knowledge concerning its venom composition. Most of Ts venom studies aim to isolate and characterize their neurotoxins, which are small, basic proteins and are eluted with high buffer concentrations on cation exchange chromatography. The first and largest fraction from carboxymethyl cellulose-52 (CMC-52) chromatography of Ts venom, named fraction I (Fr I), is a mixture of proteins of high and low molecular masses, which do not interact with the cation exchange resin, being therefore a probable source of components still unknown of this venom. Thus, the present study aimed to perform the proteome study of Fraction I from Ts venom, by high resolution mass spectrometry, and its biochemical characterization, by the determination of several enzymatic activities. Methods: Fraction I was obtained by a cation exchange chromatography using 50 mg of crude venom. This fraction was subjected to a biochemical characterization, including determination of L-amino acid oxidase, phospholipase, hyaluronidase, proteases activities and inhibition of angiotensin converting enzyme (ACE) activity. Fraction I was submitted to reduction, alkylation and digestion processes, and the tryptic digested peptides obtained were analyzed in a Q-Exactive Orbitrap mass spectrometer. Data analysis was performed by PEAKS 8.5 software against NCBI database. Results: Fraction I exhibits proteolytic activity and it was able to inhibit ACE activity. Its proteome analysis identified 8 different classes of venom components, among them: neurotoxins (48%), metalloproteinases (21%), hypotensive peptides (11%), cysteine-rich venom protein (9%), antimicrobial peptides (AMP), phospholipases and other enzymes (chymotrypsin and lysozymes) (3%) and phosphodiesterases (2%). Conclusions: The combination of a proteomic and biochemical characterization strategies leads us to identify new components in the T. serrulatus scorpion venom. The proteome of venom's fraction can provide valuable direction in the obtainment of components in their native forms in order to perform a preliminary characterization and, consequently, to promote advances in biological discoveries in toxinology. (AU)

• FAPESP's process: 14/07824-1 - Structural and functional characterization of toxins with biotechnological interest present in fraction I from Tityus serrulatus venom
• Grantee: Heloisa Tavoni Longhim Peccin
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 15/17466-8 - Venomics Platform: Pachycondyla villosa venom proteome and integrated analysis of omics data
• Grantee: Camila Takeno Cologna
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 12/14996-8 - Cloning and expression of animal toxins of biotechnological interest
• Grantee: Eliane Candiani Arantes Braga
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/26083-0 - Analysis of post-translational modifications of native and recombinant toxins from Tityus serrulatus venom by mass spectrometry
• Grantee: Fernanda Gobbi Amorim
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• 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: 14/22959-0 - Subproteome of fraction I from the venom of Tityus serrulatus scorpion by mass spectrometry
• Grantee: Heloisa Tavoni Longhim Peccin
• Support Opportunities: Scholarships abroad - Research Internship - Scientific Initiation
• FAPESP's process: 13/26200-6 - Venomics Plataform of Pachycondyla villosa Proteome, Transcriptome, Cloning and Expression of toxins with biotechnological potential
• Grantee: Camila Takeno Cologna
• Support Opportunities: Scholarships in Brazil - Post-Doctoral