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| 原矛头蝮蛇毒中一个新颖双链纤溶酶的分离纯化与性质研究 | |
| 引用本文： | 季仁升,孙黔云,乙引. 原矛头蝮蛇毒中一个新颖双链纤溶酶的分离纯化与性质研究[J]. 中国药理学通报, 2011, 27(6): 818-823. DOI: 10.3969/j.issn.1001-1978.2011.06.019 |
| 作者姓名： | 季仁升 孙黔云 乙引 |
| 作者单位： | 1. 贵州师范大学生命科学学院,贵州,贵阳,550001;贵州省中国科学院天然产物化学重点实验室,贵州,贵阳,550002 2. 贵州省中国科学院天然产物化学重点实验室,贵州,贵阳,550002 3. 贵州师范大学生命科学学院,贵州,贵阳,550001 |
| 基金项目： | 国家自然科学基金资助项目，贵州省科技计划资助项目(No黔科合SY，贵州省优秀青年科技人才资助项目 |
| 摘 要： | 目的从原矛头蝮蛇毒中寻找新型纤溶酶,对其理化性质与生物学活性进行研究,以了解其在蛇伤中的作用与毒性机制,并评估其潜在的应用价值。方法采用蛋白层析技术分离纯化获得目标蛋白,检测其分子量、等电点、肽指纹图谱、多种蛋白水解活性、抗补体活性、出血活性、水肿活性及对流血时间的影响。结果通过阴离子交换层析、凝胶过滤层析、亲和层析从原矛头蝮蛇毒中纯化出一个酸性纤溶酶PMSP-A,它是由两条等电点分别为5.7与6.1的非均等肽链共价结合而成。SDS-PAGE和凝胶过滤测定其分子量分别为26.1 ku与25.3 ku。肽指纹图谱分析表明PMSP-A与黄绿烙铁头蛇毒中的血液凝固结合因子有部分序列吻合。PMSP-A能够依次降解纤维蛋白原的Bβ、Aα链,该活性能被PMSF、1,10-phenanthroline抑制,EDTA、EGTA、SBTI不能抑制其活性。PMSP-A具有纤维蛋白、精氨酸酯水解活性,没有偶氮酪蛋白水解活性。它还具有抗补体活性。动物实验表明,PMSP-A明显延长小鼠尾静脉流血时间,能诱导小鼠足趾轻度水肿,无皮下出血活性。结论 PMSP-A是一个新颖的原矛头蝮蛇毒双链丝氨酸蛋白酶,具有多种影响机体的生物学活性。 |
| 关 键 词： | 蛇毒丝氨酸蛋白酶 纤溶酶 抗凝活性 纤维蛋白(原)水解活性 抗补体活性 蛇伤 蛇毒 原矛头蝮蛇 烙铁头蛇 |
Purification and characterization of a novel double-stranded serine protease from Protobothrops mucrosquamatus venom |
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| JI Ren-sheng,SUN Qian-yun,YI Yin. Purification and characterization of a novel double-stranded serine protease from Protobothrops mucrosquamatus venom[J]. Chinese Pharmacological Bulletin, 2011, 27(6): 818-823. DOI: 10.3969/j.issn.1001-1978.2011.06.019 | |
| Authors: | JI Ren-sheng SUN Qian-yun YI Yin |
| Affiliation: | JI Ren-sheng1,2,SUN Qian-yun2,YI Yin1(1.Life Sciences College,Guizhou Normal University,Guiyang 550001,China,2.The Key Laboratory of Chemistry for Natural Products,Guizhou Province and Chinese Academy of Sciences,Guiyang 550002,China) |
| Abstract: | Aim To discover the new or novel fibrinolysin from Protobothrops mucrosquamatus venom.The physico-chemical property and biological activity of the interest enzyme were investigated for better understanding of its role in snakebite and evaluating its clinical potential.Methods The object protein was isolated by chromotography techniques.Assays were performed to measure molecular mass,isoelectric point,peptide mass fingerprinting analysis,variety of proteolytic activity,anticomplementary activity,edema-induci... |
| Keywords: | snake venom serine proteinases fibrinolysin anticoagulant activity fibrin(ogen)olytic activity anticomplementary activity snake bite snake venom Protobothrops mucrosquamatus Trimeresurus mucrosquamatus |
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