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无机磷和叠氮钠对线粒体F1-ATPase构象的不同影响
The Different Effects of Inorganic Phosphate and Azide on the Conformation of F 1 ATPase
【摘要】 用εADP作荧光探针,用荧光猝灭的方法证明无机磷(Pi)能影响F1-ATPase的构象从而使酶结合ADP的能力降低.用aurovertinB作荧光探针研究了Pi和叠氮钠对F1-ATPase构象的影响,发现Pi能影响洗去催化位点核苷酸的F1的构象,而叠氮钠对其没有影响,但却能影响结合有ADP的F1的构象.因此Pi和叠氮钠对酶构象的影响是不同的,二者可能有不同的作用位点.
【Abstract】 H + ATPase plays a key role in oxidative phosphorylation.The soluble part F 1 ATPase was purified from pig heart mitochondria.ADP that bound to F 1 ATPase was washed off by ammonium sulfate precipitation.Using fluorescence quenching method with εADP as a fluorescence probe and acrylamide as the quencher,it is found that P i affected the conformation of F 1 ATPase and loosened ADP’s binding to F 1.The effects of P i and azide on the conformation of F 1 ATPase were also studied using aurovertin B as the fluorescence probe.The time course scanning of the fluorescence intensity of Aurovertin B indicated that P i affected the conformation of washed F 1,but azide did not.Yet azide affected the conformation of F 1·ADP complex.Thus,P i and azide have different effects on the conformation of F 1 ATPase,and may have different binding sites.
【Key words】 F 1 ATPase; Conformation; Fluorescence quenching; P i; Azide;
- 【文献出处】 中国生物化学与分子生物学报 ,CHINESE JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY , 编辑部邮箱 ,1999年03期
- 【分类号】Q554
- 【被引频次】2
- 【下载频次】51