A New Platelet-Aggregation-Inhibiting Factor Isolated from Bothrops moojeni Snake Venom.

Bruna Barbosa De Sousa,Carla Cristine Neves Mamede,Noelio Oliveira Dantas,Fábio De Oliveira,Anielle Christine Almeida Silva,Mayara Ribeiro De Queiroz,Edigar Henrique Vaz Dias,Déborah Fernanda Da Cunha Pereira,Júnia De Oliveira Costa,Mariana Santos Matias

doi:10.1155/2017/4315832

Abstract

This work reports the purification and functional characterization of BmooPAi, a platelet-aggregation-inhibiting factor from Bothrops moojeni snake venom. The toxin was purified by a combination of three chromatographic steps (ion-exchange on DEAE-Sephacel, molecular exclusion on Sephadex G-75, and affinity chromatography on HiTrap™ Heparin HP). BmooPAi was found to be a single-chain protein with an apparent molecular mass of 32 kDa on 14% SDS-PAGE, under reducing conditions. Sequencing of BmooPAi by Edman degradation revealed the amino acid sequence LGPDIVPPNELLEVM. The toxin was devoid of proteolytic, haemorrhagic, defibrinating, or coagulant activities and induced no significant oedema or hyperalgesia. BmooPAi showed a rather specific inhibitory effect on ristocetin-induced platelet aggregation in human platelet-rich plasma, whereas it had little or no effect on platelet aggregation induced by collagen and adenosine diphosphate. The results presented in this work suggest that BmooPAi is a toxin comprised of disintegrin-like and cysteine-rich domains, originating from autolysis/proteolysis of PIII SVMPs from B. moojeni snake venom. This toxin may be of medical interest because it is a platelet aggregation inhibitor, which could potentially be developed as a novel therapeutic agent to prevent and/or treat patients with thrombotic disorders.

Highlights

Snake venoms comprise pharmacologically active proteins and peptides, both enzymatic and nonenzymatic, such as phospholipases A2, metalloproteinases, serine proteinases, nucleotidases, L-amino acid oxidase, disintegrins, and Ctype lectins [1,2,3,4]
We describe the isolation and functional characterization of BmooPAi, a toxin comprised of the dis-cys domain, originating from autolysis/proteolysis of PIII snake venom metalloproteinases (SVMPs) from Bothrops moojeni snake venom, which showed an inhibitory effect on platelet aggregation
Most of these proteins belong to the SVMP family, and their effects are probably due to the presence of dis-cys domains which interact with specific protein molecules on platelet membrane surfaces [25,26,27,28,29]

Summary

Introduction

Snake venoms comprise pharmacologically active proteins and peptides, both enzymatic and nonenzymatic, such as phospholipases A2, metalloproteinases, serine proteinases, nucleotidases, L-amino acid oxidase, disintegrins, and Ctype lectins [1,2,3,4]. Several snake venom metalloproteinases (SVMPs) have been isolated and characterized by their biological activities. These enzymes play a key role in the prominent local tissue damage and systemic alterations caused by snake venom. SVMPs induce haemorrhage, myonecrosis, skin damage, inflammation, and degradation of extracellular matrix components. SVMPs comprise a group of zinc-dependent enzymes of varying molecular mass, widely distributed in Viperidae venoms. They are synthesized as multidomain precursors and stored in the venom gland as inactive zymogens [7, 9,10,11].

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Results

Conclusion