| 蝮蛇毒蛋白C激活物的纯化与活性分析 |
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| 引用本文： | 章俊,肖小璞,李长清,王宗奎,林方昭.蝮蛇毒蛋白C激活物的纯化与活性分析[J].中国输血杂志,2011,24(4):299-303. |
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| 作者姓名： | 章俊 肖小璞 李长清 王宗奎 林方昭 |
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| 作者单位： | 中国医学科学院,北京协和医学院输血研究所,四川,成都,610052 |
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| 基金项目： | 四川省科技支撑项目(2008SZ0013) |
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| 摘 要： | 目的探索从国产蝮蛇毒中分离纯化人血浆蛋白C激活物(PCA)的方法及其用于蛋白C的鉴定。方法以国产蝮蛇毒为原料,通过溶解、离心、透析,利用Q Sepharose Fast Flow离子交换层析和Superdex G75凝胶过滤法分离纯化PCA组份。通过SDS-PAGE观察PCA组份的分子量,用发色底物法和凝固法分别测定活化PC(APC)的酰胺酶活性和抗凝活性。结果经两步层析,从蝮蛇粗蛇毒中纯化出PCA组份,SDS-PAGE图谱显示在分子量51 kD左右呈现1条蛋白带;经生物学活性鉴定:该组份在体外能迅速将PC激活为APC,APC的酰胺酶活性(A405)最高约为Protac(唯一商品化的PC激活剂产品)作用的2.8倍,抗凝活性(秒值)最高约为Protac作用的1.4倍,比活性2.20 IU/mg;不同浓度组份激活PC的酰胺酶活性和抗凝活性均随组份浓度的增大而升高,有明显的量效关系。结论采用离子交换层析和凝胶过滤法,可以从国产蝮蛇粗蛇毒中分离纯化出PCA组份,该组份具有较强的激活PC的能力,其生物活性与浓度呈正相关。
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| 关 键 词： | 蝮蛇毒 纯化 活性 蛋白C特异性激活剂 PCA组份 |
Purification and activity analysis of protein C activator from Agkistrodon halys Palls |
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| ZHANG Jun,XIAO XiaoPu,LI Changqing,WANG Zhongkui,LI Fangzhao.Purification and activity analysis of protein C activator from Agkistrodon halys Palls[J].Chinese Journal of Blood Transfusion,2011,24(4):299-303. |
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| Authors: | ZHANG Jun XIAO XiaoPu LI Changqing WANG Zhongkui LI Fangzhao |
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| Institution: | ZHANG Jun,XIAO XiaoPu,LI Fangzhao,et al.Institute of Blood Transfusion,Chinese Academy of Medical Sciences & Peking Union Medical College,Chengdu 610052,China |
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| Abstract: | Objective To study the purification and biological activity of protein C activator(PCA)separated from Agkistrodon halys Palls for identification of protein C(PC).Methods Agkistrodon halys Palls was purified by ion exchange chromatography and gel filtration.The molecular weight wasere detected by SDS-PAGE.PC activity was determined with chromogenic substrate assay and one-stage clotting assay.Results Using chromatography purification,PCA was separated from Agkistrodon halys Palls,which showed a 51 kD single ... |
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| Keywords: | Agkistrodon halys Palls Purification Activity Protein C specific ity activator PCA component |
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