肖纲,刘俊琦,夏雨,张志阳,孙志良.湖南永州尖吻蝮蛇蛇毒的蛋白组分及毒性分析[J].湖南农业大学学报:自然科学版,2019,45(5):.
湖南永州尖吻蝮蛇蛇毒的蛋白组分及毒性分析
  
DOI:
中文关键词:  尖吻蝮蛇蛇毒  SDS–PAGE  Nano–LC–ESI–MS/MS  蛇毒金属蛋白  半数致死量  湖南永州
英文关键词:Deinagkistrodon acutus venom  SDS-PAGE  Nano-LC-ESI-MS/MS  Snake venom metalloproteinase  LD50  Yongzhou of Hunan
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作者单位
肖纲,刘俊琦,夏雨,张志阳,孙志良 1.湖南农业大学动物医学院湖南 长沙 4101282.湖南省兽药工程技术研究中心湖南 长沙 410128 
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中文摘要:
      采用Nano–LC–ESI–MS/MS蛋白鉴定技术对湖南永州尖吻蝮蛇蛇毒蛋白组分进行质谱分析,应用常规体外试管法对尖吻蝮蛇蛇毒体外溶血值进行检测,采用改良寇式法对尖吻蝮蛇蛇毒LD50进行测定。结果显示:尖吻蝮蛇蛇毒含有50种蛋白,蛋白相对分子质量集中在1.0×104~2.0×104(46.9%)、4.0×104~5.0×104(22.4%)、2.0×104~3.0×104(10.6%)和7.0×104~8.0×104(7.6%),其中代表性的高丰度蛋白有蛇毒金属蛋白酶A(11.7%)、蛇毒金属蛋白酶H1(9.8%)、蕲蛇类凝血酶–2(7.3%)、抗凝血酶A–A亚基(6.8%),低丰度蛋白(<0.1%)有Ecto–5'–核苷酸酶、碱性磷脂酶A2 DAV–N6、生长分化因子11;蛇毒引起红细胞溶血的最低质量浓度为60.00 μg/mL,尖吻蝮蛇蛇毒对KM小鼠的LD50为7.179 6 mg/kg,攻毒小鼠出现呼吸急促、躁动不安、注射部位出现奇特瘙痒和大面积溃烂,至死亡。
英文摘要:
      The protein components of the venom obtained from Deinagkistrodon acutus in Yongzhou area of Hunan were identified and characterized using Nano-LC-ESI-MS/MS. The in vitro hemolysis of D. acutus venom was conducted with use of the conventional test tube method, and the LD50 of the venom was measured using a challenge test in a KM mouse model. Fifty types of proteins were identified, and the protein relative molecular mass are mainly distributed in 1.0×104~ 2.0×104(46.9%), 4.0×104~5.0×104(22.4%), 2.0×104~3.0×104(10.6%) and 7.0×104~8.0×104(7.6%) containing high-abundance of representative proteins including snake venom metalloproteinase acutolysin-A(11.7%), snake venom metalloproteinase H1(9.8%), thrombin-like enzyme acutobin-2(7.3%), antithrombin A subunit A(6.8%) and low-abundance proteins(<0.1%) containing ecto-5'-nucleotidase, basic phospholipase A2 DAV-N6, growth/ differentiation factor 11. The minimal erythrocyte hemolysis concentration and LD50 of D. acutus venom were 60.00 μg/mL and 7.179 6 mg/kg, respectively. The typical clinical symptoms of challenged mice showed to be tachypnea, restlessness, unusual pruritus and widespread canker near the injection site and even to be repeatedly scratched, and death.
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