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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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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]
Total Authors: 7
[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
Total Affiliations: 3
Document type: Journal article
Source: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 25, APR 18 2019.
Web of Science Citations: 1

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: 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis
Grantee:Suely Vilela
Support type: 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 type: 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 type: Scholarships in Brazil - Post-Doctorate
Grantee:Eliane Candiani Arantes Braga
Support type: 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 type: Scholarships abroad - Research Internship - Doctorate
FAPESP's process: 15/17466-8 - Venomics Platform: Pachycondyla villosa venom proteome and integrated analysis of omics data
Grantee:Camila Takeno Cologna
Support type: Scholarships abroad - Research Internship - Post-doctor
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 type: Scholarships in Brazil - Scientific Initiation