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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2017.tde-24052017-092922
Document
Author
Full name
Caroline Serino Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2017
Supervisor
Committee
Azevedo, Anita Mitico Tanaka (President)
Oguiura, Nancy
Sant'Anna, Savio Stefanini
Tashima, Alexandre Keiji
Title in Portuguese
Purificação e caracterização do primeiro inibidor de fosfolipase A2 do tipo gama presente no soro da serpente  Bothrops jararaca.
Keywords in Portuguese
Bothrops jararaca
Biotecnologia
Fosfolipase A2
Inibidores
Soro
Abstract in Portuguese
As Fosfolipases A2 (PLA2) são enzimas que atuam desconstruindo membranas celulares, resultando em ácidos graxos e lisofosfolipidios, causando inflamação tecidual. Evidências indicam que serpentes possuem uma resistência natural devido a propriedades presentes no sangue, que inibem ações de proteínas presentes no veneno. Portanto, no presente trabalho foi isolado e caracterizado bioquimicamente e biologicamente o primeiro inibidor de PLA2 do tipo gama (γPLI) do soro da serpente B. jararaca, denominado PLI_BJ. O inibidor de PLA2 foi isolado utilizando dois passos cromatográficos. O PLI_BJ mostrou, por SDS-PAGE, uma massa molecular aparente de 25 000 e 20 000 em condições redutoras e não redutoras, respectivamente. A sequência de aminoácidos parcial de PLI_BJ foi determinada por espectrometria de massa e corresponde a 72% e 68% de cobertura da sequência de aminoácidos de duas proteínas já descritas como PLI. O PLI_BJ mostrou também atividade inibitória satisfatória nos três testes realizados sugerindo um papel deste inibidor nos efeitos de envenenamento da serpente.
Title in English
Purification and characterization of the first gamma-type phospholipase A2 inhibitor present in Bothrops jararaca snake serum.
Keywords in English
Bothrops jararaca
Biotechnology
Inhibitors
Phospholipase A2
Serum
Abstract in English
Phospholipases A2 (PLA2) are enzymes that act on cell membrane phospholipids resulting in fatty acids and lysophospholipids, deconstructing the cell wall causing tissue inflammation. Evidence indicates that snakes have natural resistance due to protective properties of blood that inhibits the action of proteins present in the venom. This study aimed to purify and characterize PLA2 inhibitors (PLI) from serum of the Bothrops jararaca snakes. PLA2 inhibitor was isolated using two chromatographic steps, and was named PLI_BJ. The purity of the PLI_BJ was confirmed by HPLC and SEC. The PLI_BJ showed, by SDS-PAGE, an molecular mass of 25,000 and 20,000 under reducing and non-reducing conditions, respectively. The partial amino acid sequence of PLI_BJ was determined by mass spectrometry and it corresponds to 72% and 68% of coverage of the amino acid sequence of two proteins already described as PLI. The PLI_BJ also showed satisfactory inhibitory activity in the three tests performed suggesting a role of this inhibitor in snake envenomation effects.
 
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Publishing Date
2017-05-24
 
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