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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2014.tde-05122014-152052
Document
Author
Full name
Bruno Duzzi
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Rioli, Vanessa (President)
Azevedo, Inácio de Loiola Meirelles Junqueira de
Oliveira, Vitor Marcelo Silveira Bueno Brandão de
Title in Portuguese
Busca de novos substratos e/ou inibidores das enzimas timet oligopeptidase (E.C.3.4.24.15) e neurolisina (E.C.3.4.24.16) nas frações de baixa massa molecular do veneno do escorpião Tityus serrulatus.
Keywords in Portuguese
Tityus serrulatus
Componentes do veneno
EP 24.15
EP 24.16
Escorpião amarelo
Oligopeptidases
Peptídeos
Abstract in Portuguese
O escorpião Tityus serrulatus é o responsável pelos acidentes mais graves no Brasil. Dentre os componentes já estudados de sua peçonha estão neurotoxinas capazes de interagir com canais iônicos, além de peptídeos biologicamente ativos também estarem presentes. Neste estudo foram isolados peptídeos da fração de baixa massa molecular da peçonha que interagiram com as oligopeptidases timet oligopeptidase (EP24.15) e neurolisina (EP24.16) através do emprego de substratos fluorescentes específicos como ferramentas. Usando espectrometria de massas, as sequências KEILG, FTR, YLPT e do análogo KELLG foram determinadas e posteriormente sintetizadas. In vitro, os peptídeos não foram substratos para enzimas já citadas, além de testes com a neprelisina e ECA. Em relação à inibição, os destaques ficam por conta de KELLG e KEILG, capazes de inibirem a EP 24.15 e de não inibirem a EP 24.16. Outro destaque foi o YLPT, apresentando um Ki de 0,94 mM perante a neprilisina. In vivo os peptídeos foram testados em relação à nocicepção, rolamento de leucócitos e reatividade vascular, onde se destacou o FTR, apresentando efeito antinociceptivo e o KEILG, capaz de aumentar o número de leucócitos, ressaltando a importância de pequenas moléculas na composição da peçonha.
Title in English
Search for new substrates and/or inhibitors of thimet oligopeptidase (EC3.4.24.15) and neurolysin (EC3.4.24.16) enzymes in low molecular weight fractions of Tityus serrulatus scorpion venom.
Keywords in English
Tityus serrulatus
EP 24.15
EP 24.16
Oligopeptidases
Peptides
Venom componentes
Yellow scorpion
Abstract in English
The scorpion Tityus serrulatus is responsible for the most serious accidents in Brazil. Among the components already studied in its venom, neurotoxins are able to interact with ion channels, and peptides with biological activity are also present. In this study, peptides from low molecular weight fraction of the venom that interacted with the oligopeptidases thimet oligopeptidase (EP24.15) and neurolysin (EP24.16) were isolated using specific fluorescent substrates as a tool. Sequences from the peptides KEILG, FTR, YLPT and the analogue KELLG were determined by mass spectrometry and subsequently synthesized. In vitro, the peptides were not substrates for those enzymes, neither for neprilysin and ACE enzymes. Concerning inhibition, the highlights are the KELLG and KEILG, which were able to inhibit EP 24.15 but not EP 24.16. Another peptide thats worth to mention is YLPT, with a Ki of 0.94 mM for neprilysin. In vivo, the peptides were tested for nociception, rolling of leukocytes and vascular reactivity, being the FTR distinguished to be able to cause antinociceptive effect and KEILG to increase the number of leukocytes. These results emphasize the importance of small molecules in the scorpion venom constitution.
 
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Publishing Date
2014-12-10
 
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