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．
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.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1999年第3期471-474,共4页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金
北京大学蛋白质工程及植物基因工程国家重点实验室基金
