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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Journal of Venomous Animals and Toxins including Tropical Diseases; v. 25, APR 18 2019.
• Citações Web of Science: 1

Resumo

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)

• Processo FAPESP: 14/07824-1 - Caracterização estrutural e funcional de toxinas de interesse biotecnológico presentes na fração I da peçonha de Tityus serrulatus
• Beneficiário: Heloisa Tavoni Longhim Peccin
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 15/17466-8 - Plataforma Venômica: Proteoma da peçonha da formiga Pachycondyla villosa e análise integrada dos resultados obtidos através de ferramentas ômicas
• Beneficiário: Camila Takeno Cologna
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 12/14996-8 - Clonagem e expressão de toxinas animais de interesse biotecnológico
• Beneficiário: Eliane Candiani Arantes Braga
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 13/26083-0 - Análise de modificações pós-traducionais de toxinas nativas e recombinantes da peçonha Tityus serrulatus por espectrometria de massas
• Beneficiário: Fernanda Gobbi Amorim
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 11/23236-4 - Toxinas animais nativas e recombinantes: análise funcional, estrutural e molecular
• Beneficiário: Suely Vilela
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 14/22959-0 - Subproteoma da fração I da peçonha do escorpião Tityus serrulatus por espectrometria de massas
• Beneficiário: Heloisa Tavoni Longhim Peccin
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Iniciação Científica
• Processo FAPESP: 13/26200-6 - Plataforma Venômica: Proteoma, Transcriptoma, Clonagem e Expressão de toxinas de interesse biotecnológico presentes na peçonha de Pachycondyla villosa
• Beneficiário: Camila Takeno Cologna
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado