Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil.
Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), Belo Horizonte, MG, Brazil.
Biomed Res Int. 2017;2017:4315832. doi: 10.1155/2017/4315832. Epub 2017 Nov 1.
This work reports the purification and functional characterization of BmooPAi, a platelet-aggregation-inhibiting factor from 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 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.
本工作报道了一种血小板聚集抑制因子 BmooPAi 的纯化和功能特征，该因子来自蛇毒。该毒素通过三种色谱步骤（DEAE-Sephacel 离子交换、Sephadex G-75 分子排阻和 HiTrap™ Heparin HP 亲和层析）的组合进行纯化。BmooPAi 在 14% SDS-PAGE 下，在还原条件下被发现为具有 32 kDa 表观分子量的单链蛋白。BmooPAi 的序列通过 Edman 降解测序揭示了 LGPDIVPPNELLEVM 的氨基酸序列。该毒素缺乏蛋白水解、出血、解聚或凝血活性，并且不会引起明显的水肿或痛觉过敏。BmooPAi 对人血小板富血浆中的瑞斯托霉素诱导的血小板聚集表现出相当特异的抑制作用，而对胶原和二磷酸腺苷诱导的血小板聚集几乎没有影响。本工作中提出的结果表明，BmooPAi 是一种由蛇毒的 PIII SVMP 自溶/蛋白水解产生的具有解体素样和富含半胱氨酸结构域的毒素。这种毒素可能具有医学意义，因为它是一种血小板聚集抑制剂，有可能开发为预防和/或治疗血栓形成障碍患者的新型治疗剂。
