眼镜王蛇（食蛇蝮属）血清中的抗出血毒素：纯化与特性鉴定

Antihemorrhagin in the blood serum of king cobra (Ophiophagus hannah): purification and characterization.

作者信息

Chanhome Lawan, Khow Orawan, Omori-Satoh Tamotsu, Sitprija Visith

机构信息

Queen Saovabha Memorial Institute, Thai Red Cross Society, 1871 Rama IV Rd., 10330, Bangkok, Thailand

出版信息

Toxicon. 2003 Jun;41(8):1013-9. doi: 10.1016/s0041-0101(03)00075-8.

DOI:10.1016/s0041-0101(03)00075-8

PMID:12875876

Abstract

King cobra (Ophiophagus hannah) serum was found to possess antihemorrhagic activity against king cobra hemorrhagin. The activity was stronger than that in commercial king cobra antivenom. An antihemorrhagin has been purified by ion exchange chromatography, affinity chromatography and gel filtration with a 22-fold purification and an overall yield of 12% of the total antihemorrhagic activity contained in crude serum. The purified antihemorrhagin was homogeneous in disc-PAGE and SDS-PAGE. Its apparent molecular weight determined by SDS-PAGE was 120 kDa. The antihemorrhagin was also active against other hemorrhagic snake venoms obtained in Thailand and Japan such as Calloselasma rhodostoma, Trimeresurus albolabris, Trimeresurus macrops and Trimeresurus flavoviridis (Japanese Habu). It inhibited the proteolytic activity of king cobra venom. It is an acid- and thermolabile protein and does not form precipitin lines against king cobra venom.

摘要

眼镜王蛇（Ophiophagus hannah）血清被发现对眼镜王蛇出血毒素具有抗出血活性。该活性比市售眼镜王蛇抗蛇毒血清中的活性更强。一种抗出血毒素已通过离子交换色谱、亲和色谱和凝胶过滤进行纯化，纯化倍数为22倍，粗血清中总抗出血活性的总产率为12%。纯化后的抗出血毒素在圆盘聚丙烯酰胺凝胶电泳（disc-PAGE）和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）中均呈均一性。通过SDS-PAGE测定其表观分子量为120 kDa。该抗出血毒素对在泰国和日本获得的其他出血性蛇毒也有活性，如圆斑蝰（Calloselasma rhodostoma）、白唇竹叶青（Trimeresurus albolabris）、大眼竹叶青（Trimeresurus macrops）和日本蝮蛇（Trimeresurus flavoviridis，日本哈布蛇）。它抑制眼镜王蛇毒的蛋白水解活性。它是一种对酸和热不稳定的蛋白质，与眼镜王蛇毒不形成沉淀线。