Isolation and functional characterization of proinflammatory acidic phospholipase A₂ from Bothrops leucurus snake venom

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• 作者：Dé ; bora C.O. Nunes^a ; Renata S. Rodrigues^b ; Malson N. Lucena^c ; Camila T. Cologna^b ; Ana Carolina S. Oliveira^d ; Amé ; lia Hamaguchi^a ; Maria I. Homsi-Brandeburgo^a ; Eliane C. Arantes^b ; David N.S. Teixeira^d ; Carlos Ueira-Vieira^a ; Veridiana M. Rodrigues^a ; ^e ; ^{veridiana@ingeb.ufu.br"" rel=""nofollow}
• 关键词：Acidic phospholipase A₂ ; Bothrops leucurus ; Cytokines ; Proinflammatory profile ; Snake venom
• 刊名：Comparative Biochemistry and Physiology, Part C
• 出版年：2011
• 出版时间：September 2011
• 年：2011
• 卷：154
• 期：3
• 页码：226-233
• 全文大小：893 K

文摘

In the present study, an acidic PLA₂, designated Bl-PLA₂, was isolated from Bothrops leucurus snake venom through two chromatographic steps: ion-exchange on CM-Sepharose and hydrophobic chromatography on Phenyl-Sepharose. Bl-PLA₂ was homogeneous on SDS-PAGE and when submitted to 2D electrophoresis the molecular mass was 15,000 Da and pI was 5.4. Its N-terminal sequence revealed a high homology with other Asp49 acidic PLA₂s from snake venoms. Its specific activity was 159.9 U/mg and the indirect hemolytic activity was also higher than that of the crude venom. Bl-PLA₂ induced low myotoxic and edema activities as compared to those of the crude venom. Moreover, the enzyme was able to induce increments in IL-12p40, TNF-α, IL-1β and IL-6 levels and no variation of IL-8 and IL-10 in human PBMC stimulated in vitro, suggesting that Bl-PLA₂ induces proinflammatory cytokine production by human mononuclear cells. Bothrops leucurus venom is still not extensively explored and knowledge of its components will contribute for a better understanding of its action mechanism.