绿豆下胚轴质膜微囊的提取和H^+-ATPase活性研究  

Isolation of Mung Bean Plasma Membrane Vesicles and the Analyses of Hydrolysis Activity of PM H^+-ATPase

作　　者：文彬[1] 宾金华[1] 潘瑞炽[1] 王小菁[1] 

机构地区：[1]华南师范大学生命科学学院广东省植物发育生物工程重点实验室,广州510631

出　　处：《植物学通报》2004年第3期319-325,共7页Chinese Bulletin of Botany

基　　金：教育部留学回国人员科研启动基金资助;优秀年轻教师基金资助;国家自然科学基金(39970079)资助。

摘　　要：以两相法提取纯化绿豆下胚轴质膜微囊,材料与两相体系重量之比为32∶8时,一次洗膜就可以得到纯度较高的质膜微囊。提取缓冲液中牛血清白蛋白的浓度对质膜H+-ATPase的潜在活性有影响。质膜H+-ATPase水解活性依赖于Mg2+,Ca2+对酶活性有明显的促进作用。壳梭孢素(fusicoccin, FC)对酶有明显的刺激作用,活体条件最大刺激达到72%,而离体条件下刺激为30%。The aqueous two-phase partition method was used to isolate and purify plasma membrane(PM) vesicles from mung bean hypocotyls. The PM vesicles were purified well when the proportionof the weight of hypocotyls and two-phase partition reached 32:8. Bovine serum albumin in homog-enization medium influenced the latency activity of the PM H+-ATPase. The enzyme hydrolysisactivity was Mg2+ dependent and was strongly stimulated byCa2+. Fusicoccin (FC), an activator ofPM H+-ATPase, stimulated the enzyme activity with the maximal increase of 72% in vivo and of 30%in vitro as compared with control.

关 键 词：绿豆 下胚轴 质膜微囊 牛血清白蛋白 缓冲液 

分 类 号：S522[农业科学—作物学]