液泡膜H^+-ATPase质子泵活性的荧光测定  

Fluorescence Measurement of Proton Pumping of Tonoplast H + ATPase.

作　　者：王欢[1] 祝雄伟[1] 王延枝[1] 

机构地区：[1]武汉大学生命科学学院

出　　处：《生物化学与生物物理进展》1999年第2期184-187,共4页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金

摘　　要：使用荧光猝灭法测定植物液泡膜Ｈ＋ＡＴＰａｓｅ质子转运活性．比较了两种常用荧光染料吖啶橙和喹亚因在不同浓度的测定灵敏度．探讨了不同蛋白量和缓冲系统对测定结果的影响．得到了用５μｍｏｌ／Ｌ吖啶橙，２００～２５０μｇ蛋白质含量，ＨｅｐｅｓＴｒｉｓ（ｐＨ７０）为缓冲介质。Fluorescence quenching method was to measure the proton pumping activity of tonoplast H + ATPase from higher plants. The sensitivity of two common fluorescent dyes was compared. Acridine orange is better than quinacrine with the initial slope and maximum quenching. The optimim for the fluorescence measurement of acridine orange dye were: 5 μmol/L acridine orange, 200～250 μg protein, Hepes Tris of Hepes BTP(pH 7 0). Initiating with ATP or Mg 2+ was both valid.

关 键 词：液泡膜 H^+-ATPASE 泵活性 荧光猝灭法 

分 类 号：Q73[生物学—分子生物学] Q6－3