白唇竹叶青蛇毒5′-核苷酸酶的分离纯化及性质  

Purification and Characterization of 5′-nucleotidase from Trimeresurus albolabris Venom

作　　者：陈夏[1] 余晓东[1] 邓敏[1] 李卉[1] 林亦心[1] 和七一[1] 柳建平[1] 

机构地区：[1]重庆师范大学生命科学学院,重庆市生物活性物质工程研究中心,重庆市动物生物学重点实验室,重庆400047

出　　处：《Zoological Research》2008年第4期399-404,共6页动物学研究（英文）

基　　金：重庆市自然科学基金重点项目(CSTC2006BA5032);重庆市教委基金项目(KJ080824)

摘　　要：用DEAE-SephadexA-25、Sephadex-G-100和CM-SephadexC-50三步柱层析分离法,从白唇竹叶青(Trimeresurus albolabris)蛇毒中分离纯化出具有5′-核苷酸酶活性的组分。SDS-聚丙烯酰胺凝胶电泳测定其分子量为48.03kDa,HPLC柱层析图谱为单一峰。该组分是一个糖蛋白,以一磷酸腺苷(AMP)为底物时,其酶活力为330.33μg Pi/(min·mg);而以二磷酸腺苷(ADP)为底物时,其酶活力为123.56μg Pi/(min·mg)。金属离子Zn2+、Fe3+和Cu2+对5′-核苷酸酶活性有显著的抑制作用,EDTA可完全抑制其酶活性。该酶的最适pH为9,最适温度为50℃。该组分还具有抑制由ADP诱导的血小板聚集的生物功能。A 5＇-nucleotidase was isolated and purified from the snake venom of T. albolabris using three steps of chromatography including DEAE-SephadexA-25, Sephadex-G-100 and CM-Sephadex C-50. Using SDS-PAGE and HPLC column chromatography the purified 5＇-nucleotidase proved to be homogenous. It was a glycoprotein with a molecular weight of 48.03kDa. The enzymatic activities of the purified 5＇-nucleotidase were 330.331ag Pi/min mg and 123.561μg Pi/min mg when using AMP （adenosine monophosphate） and ADP （adenosine diphosphate） as substrates, respectively. Metal ions, including Zn^2＋, Fe^3＋ and Cu^2＋, could inhibit 5＇-nucleotidase activity, as did EDTA. Its optimum pH was nine and its optimum temperature was 50℃ It has a potent inhibitory effect on rabbit platelet aggregation induced by ADP.

关 键 词：白唇竹叶青蛇毒 5'-核苷酸酶 分离纯化 

分 类 号：Q55[生物学—生物化学]