Tai Hong, Wei Qin, Jin Yang, Su Min, Song Jian-Xin, Zhou Xin-Ding, Ouyang Hong-Mei, Wang Wan-Yu, Xiong Yu-Liang, Zhang Yun
Department of Animal Toxinology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, Yunnan, People's Republic of China.
Toxicon. 2004 Nov;44(6):649-56. doi: 10.1016/j.toxicon.2004.07.022.
TMVA is a C-type lectin-like protein with potent platelet activating activity from Trimeresurus mucrosquamatus venom. In the absence of von Willebrand factor (vWF), TMVA dose-dependently induced aggregation of washed platelets. Anti-GP Ib monoclonal antibodies (mAbs), HIP1, specifically inhibited TMVA-induced aggregation in a dose-dependent manner. The aggregation was also inhibited by mAb P2 (an anti-GP IIb mAb). Flow cytometric analysis revealed that FITC-TMVA bound to human formalin-fixed platelets in a saturable manner, and its binding was specifically blocked by HIP1 in a dose-dependent manner. Flow cytometric analysis showed that TMVA did not bind to platelet GPIX, GPIIb, GPIIIa, GPIa, GPIIa and GPIV. Moreover, the platelet aggregation induced by TMVA was partially inhibited when platelet was pretreated with mocarhagin, a snake venom protease that specifically cleaves human GPIb. These results suggest that TMVA is a strong platelet agonist via GPIb and it might have multiple functional binding-sites on GPIb molecule or on other unknown receptor.
竹叶青毒素A是一种来自竹叶青蛇毒液的具有强大血小板激活活性的C型凝集素样蛋白。在没有血管性血友病因子(vWF)的情况下,竹叶青毒素A能剂量依赖性地诱导洗涤后的血小板聚集。抗糖蛋白Ib单克隆抗体(mAb)HIP1能以剂量依赖性方式特异性抑制竹叶青毒素A诱导的聚集。单克隆抗体P2(一种抗糖蛋白IIb单克隆抗体)也能抑制这种聚集。流式细胞术分析显示,异硫氰酸荧光素标记的竹叶青毒素A以可饱和的方式与人福尔马林固定的血小板结合,并且其结合被HIP1以剂量依赖性方式特异性阻断。流式细胞术分析表明,竹叶青毒素A不与血小板糖蛋白IX、糖蛋白IIb、糖蛋白IIIa、糖蛋白Ia、糖蛋白IIa和糖蛋白IV结合。此外,当血小板用莫卡哈金(一种特异性切割人糖蛋白Ib的蛇毒蛋白酶)预处理时,竹叶青毒素A诱导的血小板聚集会受到部分抑制。这些结果表明,竹叶青毒素A是一种通过糖蛋白Ib起作用的强效血小板激动剂,并且它可能在糖蛋白Ib分子或其他未知受体上具有多个功能性结合位点。