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Anti-platelet aggregation activity of two novel acidic Asp49-phospholipases A 2 from Bothrops brazili snake venom.

Authors :
Sobrinho JC
Kayano AM
Simões-Silva R
Alfonso JJ
Gomez AF
Gomez MCV
Zanchi FB
Moura LA
Souza VR
Fuly AL
de Oliveira E
da Silva SL
Almeida JR
Zuliani JP
Soares AM
Source :
International journal of biological macromolecules [Int J Biol Macromol] 2018 Feb; Vol. 107 (Pt A), pp. 1014-1022. Date of Electronic Publication: 2017 Sep 23.
Publication Year :
2018

Abstract

Phospholipases A <subscript>2</subscript> (PLA <subscript>2</subscript> s) are important enzymes present in snake venoms and are related to a wide spectrum of pharmacological effects, however the toxic potential and therapeutic effects of acidic isoforms have not been fully explored and understood. Due to this, the present study describes the isolation and biochemical characterization of two new acidic Asp49-PLA <subscript>2</subscript> s from Bothrops brazili snake venom, named Braziliase-I and Braziliase-II. The venom was fractionated in three chromatographic steps: ion exchange, hydrophobic interaction and reversed phase. The isoelectric point (pI) of the isolated PLA <subscript>2</subscript> s was determined by two-dimensional electrophoresis, and 5.2 and 5.3 pIs for Braziliase-I and II were observed, respectively. The molecular mass was determined with values ​​of 13,894 and 13,869Da for Braziliase-I and II, respectively. Amino acid sequence by Edman degradation and mass spectrometry completed 87% and 74% of the sequences, respectively for Braziliase-I and II. Molecular modeling of isolated PLA <subscript>2</subscript> s using acid PLA <subscript>2</subscript> BthA-I-PLA <subscript>2</subscript> from B. jararacussu template showed high quality. Both acidic PLA <subscript>2</subscript> s showed no significant myotoxic activity, however they induced significant oedematogenic activity. Braziliase-I and II (100μg/mL) showed 31.5% and 33.2% of cytotoxicity on Trypanosoma cruzi and 26.2% and 19.2% on Leishmania infantum, respectively. Braziliase-I and II (10μg) inhibited 96.98% and 87.98% of platelet aggregation induced by ADP and 66.94% and 49% induced by collagen, respectively. The acidic PLA <subscript>2</subscript> s biochemical and structural characterization can lead to a better understanding of its pharmacological effects and functional roles in snakebites pathophysiology, as well as its possible biotechnological applications as research probes and drug leads.<br /> (Copyright © 2017 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1879-0003
Volume :
107
Issue :
Pt A
Database :
MEDLINE
Journal :
International journal of biological macromolecules
Publication Type :
Academic Journal
Accession number :
28951306
Full Text :
https://doi.org/10.1016/j.ijbiomac.2017.09.069