Molecules 2014, 19(3), 3552-3569; doi:10.3390/molecules19033552
Article

Protein-Rich Fraction of Cnidoscolus urens (L.) Arthur Leaves: Enzymatic Characterization and Procoagulant and Fibrinogenolytic Activities

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Received: 20 December 2013; in revised form: 13 March 2014 / Accepted: 14 March 2014 / Published: 21 March 2014
(This article belongs to the Section Metabolites)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: Proteolytic enzymes are important macromolecules in the regulation of biochemical processes in living organisms. Additionally, these versatile biomolecules have numerous applications in the industrial segment. In this study we have characterized a protein-rich fraction of Cnidoscolus urens (L.) Arthur leaves, rich in proteolytic enzymes, and evaluated its effects on the coagulation cascade. Three protein-rich fractions were obtained from the crude extract of C. urens leaves by precipitation with acetone. Fraction F1.0 showed higher proteolytic activity upon azocasein, and thus, was chosen for subsequent tests. The proteolytic activity of F1.0 on fibrinogen was dose-dependent and time-dependent. The extract demonstrated procoagulant activity on citrated plasma and reduced the APTT, not exerting effects on PT. Despite the fibrin(ogen)olytic activity, F1.0 showed no defibrinogenating activity in vivo. The fraction F1.0 did not express hemorrhagic nor hemolytic activities. The proteolytic activity was inhibited by E-64, EDTA and in the presence of metal ions, and increased when pretreated with reducing agents, suggesting that the observed activity was mostly due to cysteine proteases. Several bands with proteolytic activity were detected by zymography with gelatin, albumin and fibrinogen. The optimal enzymatic activity was observed in temperature of 60 °C and pH 5.0, demonstrating the presence of acidic proteases. In conclusion, these results could provide basis for the pharmacological application of C. urens proteases as a new source of bioactive molecules to treat bleeding and thrombotic disorders.
Keywords: Cnidoscolus urens; Euphorbiaceae; cysteine proteases; enzymatic characterization; fibrin(ogen)olytic; procoagulant
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MDPI and ACS Style

de Menezes, Y.A.S.; Félix-Silva, J.; da Silva-Júnior, A.A.; Rebecchi, I.M.M.; de Oliveira, A.S.; Uchoa, A.F.; Fernandes-Pedrosa, M.F. Protein-Rich Fraction of Cnidoscolus urens (L.) Arthur Leaves: Enzymatic Characterization and Procoagulant and Fibrinogenolytic Activities. Molecules 2014, 19, 3552-3569.

AMA Style

de Menezes YAS, Félix-Silva J, da Silva-Júnior AA, Rebecchi IMM, de Oliveira AS, Uchoa AF, Fernandes-Pedrosa MF. Protein-Rich Fraction of Cnidoscolus urens (L.) Arthur Leaves: Enzymatic Characterization and Procoagulant and Fibrinogenolytic Activities. Molecules. 2014; 19(3):3552-3569.

Chicago/Turabian Style

de Menezes, Yamara A.S.; Félix-Silva, Juliana; da Silva-Júnior, Arnóbio A.; Rebecchi, Ivanise M.M.; de Oliveira, Adeliana S.; Uchoa, Adriana F.; Fernandes-Pedrosa, Matheus F. 2014. "Protein-Rich Fraction of Cnidoscolus urens (L.) Arthur Leaves: Enzymatic Characterization and Procoagulant and Fibrinogenolytic Activities." Molecules 19, no. 3: 3552-3569.


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