Properties and cDNA cloning of antihemorrhagic factors in sera of Chinese and Japanese mamushi (Gloydius blomhoffi)
Properties and cDNA cloning of antihemorrhagic factors in sera of Chinese and Japanese mamushi (Gloydius blomhoffi)
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DOI：
10.1016/j.toxicon.2007.09.007
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发表时间：
2008-02-01
期刊：
影响因子：
2.8
通讯作者：
Terada, Shigeyuki
中科院分区：
文献类型：
--
作者：
Aoki, Narumi;Tsutsumi, Kadzuyo;Terada, Shigeyuki
采用乙醇沉淀结合反相高效液相色谱法（HPLC），在C_8柱上从中华蝮蛇（Gloydius blomhoffi brevicaudus）血清中分离出一种抗出血蛋白。这种蛋白质命名为中国mamushi血清因子（cMSF）抑制mamushi毒液诱导的出血以剂量依赖性的方式。它对胰蛋白酶、糜蛋白酶、嗜热菌蛋白酶和木瓜蛋白酶没有影响，但抑制了几种蛇毒金属蛋白酶（SVMPs）的蛋白酶活性，包括从mamushi和habu（竹叶青蛇）毒液中分离的出血酶。从日本曼蚊（G. mamushi）血清中也纯化出具有抗出血活性的类似蛋白（Japanese MSF，jMSF）。blomhoffi）。直接分析了完整的cMSF和jMSF的N-末端70和51个残基;注意到两个MSF的序列与来自哈布血清的抗出血蛋白（HSF）的序列之间的相似性。为了获得MSFs的完整氨基酸序列，根据其N-末端氨基酸序列，从中国和日本蝰蛇的肝脏mRNA中克隆编码这些蛋白的cDNA。两种MSF的成熟形式均由305个氨基酸组成，具有19个残基的信号序列，并且在其富含His的结构域中检测到独特的17个残基缺失。(c)2007爱思唯尔有限公司保留所有权利。
An antihemorrhagic protein has been isolated from the serum of Chinese mamushi (Gloydius blomhoffi brevicaudus) by using a combination of ethanol precipitation and a reverse-phase high-performance liquid chromatography (HPLC) on a C8 column. This protein-designated Chinese mamushi serum factor (cMSF)-suppressed mamushi venom-induced hemorrhage in a dose-dependent manner. It had no effect on trypsin, chymotrypsin, thermolysin, and papain but inhibited the proteinase activities of several snake venom metalloproteinases (SVMPs) including hemorrhagic enzymes isolated from the venoms of mamushi and habu (Trimeresurus flavoviridis). A similar protein (Japanese MSF, jMSF) with antihemorrhagic activity has also been purified from the sera of Japanese mamushi (G. blomhoffi). The N-terminal 70 and 51 residues of the intact cMSF and jMSF were directly analyzed; a similarity between the sequences of two MSFs to that of antihemorrhagic protein (HSF) from habu serum was noticed. To obtain the complete amino acid sequences of MSFs, cDNAs encoding these proteins were cloned from the liver mRNA of Chinese and Japanese vipers based on their N-terminal amino acid sequences. The mature forms of both MSFs consisted of 305 amino acids with a 19-residue signal sequence, and a unique 17-residue deletion was detected in their His-rich domains. (c) 2007 Elsevier Ltd. All rights reserved.