Article (Scientific journals)
Proteome of fraction from Tityus serrulatus venom reveals new enzymes and toxins
Amorim, F. G.; Longhim, H. T.; Cologna, C. T. et al.
2019In Journal of Venomous Animals and Toxins including Tropical Diseases, 25
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Keywords :
ACE inhibitors; Enzymes; Proteases; Proteome; Scorpion venom; Tityus serrulatus; Tityus toxin; Article
Abstract :
[en] 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. © The Author(s).
Disciplines :
Chemistry
Author, co-author :
Amorim, F. G.;  Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil, University of Vila Velha, Vila Velha, ES, Brazil
Longhim, H. T.;  Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil
Cologna, C. T.;  Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil, Laboratory of Mass Spectrometry, MolSys Research Unit, Liège Université, Liège, Belgium
Degueldre, Michel ;  Université de Liège - ULiège > MolSys
De Pauw, Edwin  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Quinton, Loïc  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique
Arantes, E. C.;  Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café s/n, Monte Alegre, Ribeirão Preto, SP, 14040-903, Brazil
Language :
English
Title :
Proteome of fraction from Tityus serrulatus venom reveals new enzymes and toxins
Publication date :
2019
Journal title :
Journal of Venomous Animals and Toxins including Tropical Diseases
eISSN :
1678-9199
Publisher :
Centro de Estudos de Venenos e Animais Peconhentos
Volume :
25
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico [BR]
CAPES - Coordenação de Aperfeicoamento de Pessoal de Nível Superior [BR]
FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo [BR]
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