Leimgruber R M, Senior A E
J Biol Chem. 1976 Nov 25;251(22):7110-3.
Phosphorylating submitochondrial particles from beef heart (ETPH) prepared here contained about 2.4 nmol of ATP and 1.9 nmol of ADP/mg of protein after repeated washing of the particles. Essentially all of the "tightly bound " ATP and ADP was removed by trypsin treatment. The trypsin-treated ETPH had increased ATPase activity, undiminished NADH oxidase and succinate oxidase activity, but energy-coupling activity (ATP-driven reversed electron transfer) was abolished. Removal of half the ATP and ADP occurred at low levels of trypsin and was associated with loss of half of the coupling activity. Gel filtration of ETPH in high ionic strength buffer also removed ADP and ATP from the particles, resulting in loss of energy-coupling activity, while ATPase activity was increased. The results support the contention that the tightly bound ADP is essential in energy coupling in mitochondria. Tightly bound ATP may also play an essential role.
此处制备的来自牛心的磷酸化亚线粒体颗粒（ETPH）在反复洗涤颗粒后，每毫克蛋白质含有约2.4纳摩尔的ATP和1.9纳摩尔的ADP。基本上所有“紧密结合”的ATP和ADP都通过胰蛋白酶处理被去除。经胰蛋白酶处理的ETPH的ATP酶活性增加，NADH氧化酶和琥珀酸氧化酶活性未减弱，但能量偶联活性（ATP驱动的逆向电子传递）被消除。在低水平的胰蛋白酶作用下，一半的ATP和ADP被去除，同时伴随着一半偶联活性的丧失。在高离子强度缓冲液中对ETPH进行凝胶过滤也从颗粒中去除了ADP和ATP，导致能量偶联活性丧失，而ATP酶活性增加。这些结果支持了紧密结合的ADP在线粒体能量偶联中至关重要的观点。紧密结合的ATP可能也起着至关重要的作用。
