Isolation of a highly active H+-ATPase from beef heart mitochondria.
Isolation of a highly active H+-ATPase from beef heart mitochondria.
复制标题
从牛心线粒体中分离出高活性 H -ATP 酶。
DOI：
10.1007/bf00743058
复制
发表时间：
1982
影响因子：
3
通讯作者：
Sanadi,DR
中科院分区：
文献类型：
--
作者：
Hughes,J;Joshi,S;Torok,K;Sanadi,DR
Sadleret al. (1974) 最初描述的溶血卵磷脂提取程序经过修改，可产生具有高水平 Pi-ATP 交换活性 (400–600 nmol × min−1× mg−1) 的 H+-ATP 酶。在存在 asolectin 的情况下进行蔗糖密度梯度离心后，该活性进一步增强（1400–1600 nmol × min−1× mg−1）。这种增强部分来自脂质依赖性激活，部分来自非活性复合物的去除。使用 oxonol VI 的结合作为膜电位的指示剂，通过分光光度法测定了复合物的 H+ 转位活性。 Pi-ATP 交换、ATP 水解和 oxonol 结合对能量转移抑制剂（寡霉素、鲁他霉素）和/或解偶联剂（DNP、FCCP）敏感。
The lysolecithin extraction procedure originally described by Sadleret al.(1974) has been modified to yield a H+-ATPase with high levels of Pi-ATP exchange activity (400–600 nmol × min−1× mg−1). This activity is further enhanced (1400–1600 nmol × min−1× mg−1) following sucrose density gradient centrifugation in the presence of asolectin. This enhancement results in part from a lipid-dependent activation and in part from removal of inactive complexes. The H+translocating activity of the complex has been determined spectrophotometrically using binding of oxonol VI as an indicator of membrane potential. Pi-ATP exchange, ATP hydrolysis, and oxonol binding are sensitive to energy-transfer inhibitors (oligomycin, rutamycin) and/or uncouplers (DNP, FCCP).